Novel 4-dedimethylaminotetracycline derivatives

ABSTRACT

The present invention provides new chemically modified 4-dedimethylaminotetracycline compounds that can be substituted with aryl, alkenyl, or alkynyl groups. The 7, 8, and/or 9 positions and methods for preparing such compounds. Other tetracycline compounds include the 4-dedimethylaminotetracycline derivatives with an oxime group, NH-Alkyl, or N-NH-Alkyl group substituted at the C4 position as well as C2 Mannich derivatives. The present invention also provides a method of treating a mammal suffering from conditions or diseases by administering to the mammal an effective amount of the new chemically modified tetracycline compounds.

[0001] The present application is a continuation-in-part of Ser. No.09/573,654, filed May 18, 2000 which is a continuation-in-part of Ser.No. 09/479,604, filed Jan. 7, 2000, which is a continuation-in-part ofSer. No. 09/195,013 filed Nov. 18, 1998, all of which are incorporatedherein by reference.

FIELD OF INVENTION

[0002] The present invention relates to novel4-dedimethylaminotetracycline derivatives, methods for producing thenovel derivatives and methods of using these derivatives.

BACKGROUND OF THE INVENTION

[0003] The compound, tetracycline, exhibits the following generalstructure:

[0004] The numbering system of the ring nucleus is as follows:

[0005] Tetracycline as well as the 5-OH (Terramycin) and 7-Cl(Aureomycin) derivatives exist in nature, and are well knownantibiotics. Natural tetracyclines may be modified without losing theirantibiotic properties, although certain elements of the structure mustbe retained. The modifications that may and may not be made to the basictetracycline structure have been reviewed by Mitscher in The Chemistryof Tetracyclines, Chapter 6, Marcel Dekker, Publishers, New York (1978).According to Mitscher, the substituents at positions 5-9 of thetetracycline ring system may be modified without the complete loss ofantibiotic properties. Changes to the basic ring system or replacementof the substituents at positions 1-4 and 10-12, however, generally leadto synthetic tetracyclines with substantially less or effectively noantimicrobial activity. Some examples of chemically modifiednon-antimicrobial tetracyclines (hereinafter CMT) include4-dedimethylaminotetracyline, 4-dedimethylaminosancycline(6-demethyl-6-deoxy-4-dedimethylaminotetracycline),4-dedimethylaminominocycline(7-dimethylamino-4-dedimethylaminotetracycline), and4-dedimethylaminodoxycycline(5-hydroxy-6-deoxy-4-dedimethyaminotetracycline).

[0006] Some 4-dedimethylaminotetracyline derivatives are disclosed inU.S. Pat. Nos. 3,029,284 and 5,122,519. They include6-demethyl-6-deoxy-4-dedimethylaminotetracycline and5-hydroxy-6-deoxy-4-dedimethylaminotetracycline with hydrogen and othersubstituents at the C7, and the C9 positions on the D ring. Thesesubstituents include amino, nitro, di (lower alkyl) amino, and mono(lower alkyl) amino or halogen. The6-demethyl-6-deoxy-4-dedimethylaminotetracycline derivatives and5-hydroxy-6-deoxy-4-dedimethylaminotetracycline derivatives are said tobe useful as antimicrobial agents.

[0007] Other 4-dedimethylaminotetracycline derivatives with an oximegroup at the C4 position on the A ring are disclosed in U.S. Pat. Nos.3,622,627 and 3,824,285. These oxime derivatives have hydrogen andhalogen as substituents at the C7 position and include7-halo-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline, and7-halo-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline.

[0008] Alkylamino (NH-alkyl), and alkylhydrazone (N-NH-alkyl) groupshave been substituted on the A ring at the C4 position of4-dedimethylaminotetracycline. These compounds are known for theirantimicrobial properties. See U.S. Pat. Nos. 3,345,370, 3,609,188,3,622,627, 3,824,285, 3,622,627, 3,502,660, 3,509,184, 3,502,696,3,515,731, 3,265,732, 5,122,519, 3,849,493, 3,772,363, and 3,829,453.

[0009] In addition to their antimicrobial properties, tetracyclines havebeen described as having a number of other uses. For example,tetracyclines are also known to inhibit the activity of collagendestructive enzymes, such as matrix metalloproteinases (MMP), includingcollagenase (MMP-1), gelatinase (MMP-2) and stromelysin (MMP-3). Golubet al., J. Periodont. Res. 20:12-23 (1985); Golub et al. Crit. Revs.Oral Biol. Med. 2: 297-322 (1991); U.S. Pat. Nos. 4,666,897; 4,704,383;4,935,411; 4,935,412. Also, tetracyclines have been known to inhibitwasting and protein degradation in mammalian skeletal muscle, U.S. Pat.No. 5,045,538, and to enhance IL-10 production in mammalian cells.

[0010] Furthermore, tetracyclines were reported to enhance bone proteinsynthesis in U.S. Pat. No. Re. 34,656, and to reduce bone resorption inorgan culture in U.S. Pat. No. 4,704,383.

[0011] Similarly, U.S. Pat. No. 5,532,227 to Golub et al, discloses thattetracyclines can ameliorate the excessive glycosylation of proteins. Inparticular, tetracyclines inhibit the excessive collagen cross linkingwhich results from excessive glycosylation of collagen in diabetes.

[0012] Tetracyclines are known to inhibit excessive phospholipase A₂activity involved in inflammatory conditions such as psoriasis asdisclosed in U.S. Pat. No. 5,532,227. In addition, tetracyclines arealso known to inhibit cycloxygenase-2 (COX-2), tumor necrosis factor(TNF), nitric oxide and IL-1 (interleukin-1).

[0013] These properties cause the tetracyclines to be useful in treatinga number of diseases. For example, there have been a number ofsuggestions that tetracyclines, including non-antimicrobialtetracyclines, are effective in treating arthritis. See, for example,Greenwald, et al. “Tetracyclines Suppress Metalloproteinase Activity inAdjuvant Arthritis and, in Combination with Flurbiprofen, AmeliorateBone Damage,” Journal of Rheumatology 19:927-938(1992); Greenwald etal., “Treatment of Destructive Arthritic Disorders with MMP Inhibitors:Potential Role of Tetracyclines in Inhibition of MatrixMetalloproteinases: Therapeutic Potential,” Annals of the New YorkAcademy of Sciences 732: 181-198 (1994); Kloppenburg, et al.“Minocycline in Active Rheumatoid Arthritis,” Arthritis Rheum37:629-636(1994); Ryan et al., “Potential of Tetracycline to ModifyCartilage Breakdown in Osteoarthritis,” Current Opinion in Rheumatology8: 238-247(1996); O'Dell et al, “Treatment of Early Rheumatoid Arthritiswith Minocycline or Placebo,” Arthritis. Rheum. 40:842-848(1997).

[0014] Tetracyclines have also been suggested for use in treating skindiseases. For example, White et al., Lancet, April 29, p. 966 (1989)report that the tetracycline minocycline is effective in treatingdystrophic epidermolysis bullosa, which is a life-threatening skincondition believed to be related to excess collagenase.

[0015] Furthermore, studies have also suggested that tetracyclines andinhibitors of metalloproteinases inhibit tumor progression, DeClerck etal., Annals N.Y. Acad. Sci., 732: 222-232 (1994), bone resorption,Rifkin et al., Annals N.Y. Acad. Sci., 732: 165-180 (1994),angiogenesis, Maragoudakis et al., Br. J. Pharmacol., 111: 894-902(1994), and may have anti-inflammatory properties, Ramamurthy et al.,Annals N.Y. Acad. Sci., 732, 427-430 (1994).

[0016] Based on the foregoing, tetracyclines have been found to beeffective in treating numerous diseases and conditions. Therefore, thereis a need for new and even more useful 4-dedimethylaminotetracyclinederivatives.

SUMMARY OF THE INVENTION

[0017] It has now been discovered that these and other objectives can beachieved by tetracycline compounds of the formulae:

[0018] wherein R7 is selected from the group consisting of hydrogen,amino, nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of hydrogen, amino,azido, nitro, acylamino, hydroxy, ethoxythiocarbonylthio, mono(loweralkyl)amino, halogen, diazonium, di(lower alkyl)amino and RCH(NH₂)CO; Ris hydrogen or lower alkyl; and pharmaceutically acceptable andunacceptable salts thereof; with the following provisos: when either R7and R9 are hydrogen then R8 must be halogen; and when R6-a, R6, R5 andR9 are all hydrogen and R7 is hydrogen, amino, nitro, halogen,dimethylamino or diethylamino, then R8 must be halogen; and when R6-a ismethyl, R6 and R9 are both hydrogen, R5 is hydroxyl and R7 is hydrogen,amino, nitro, halogen or diethylamino, then R8 is halogen; and when R6-ais methyl, R6 is hydroxyl, R5, R7 and R9 are all hydrogen, then R8 mustbe halogen; and when R6-a, R6 and R5 are all hydrogen, R9 is methylaminoand R7 is dimethylamino, then R8 must be halogen; and when R6-a ismethyl, R6 is hydrogen, R5 is hydroxyl, R9 is methylamino and R7 isdimethylamino, then R8 must be halogen; and when R6-a is methyl, R6, R5and R9 are all hydrogen and R7 is cyano, then R8 must be halogen.

[0019] In another embodiment, the invention provides a tetracyclinecompound of the formulae:

[0020] wherein R7 is selected from the group consisting of hydrogen,amino, nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R4 is selected from the group consisting of NOH, N-NH-A, andNH-A, where A is a lower alkyl group; R8 is selected from the groupconsisting of hydrogen and halogen; R9 is selected from the groupconsisting of hydrogen, amino, azido, nitro, acylamino, hydroxy,ethoxythiocarbonylthio, mono(lower alkyl) amino, halogen, di(loweralkyl)amino and RCH(NH₂)CO; R is hydrogen or lower alkyl; andpharmaceutically acceptable and unacceptable salts thereof; with thefollowing provisos: when R4 is NOH, N-NH-alkyl or NH-alkyl and R7, R6-a,R6, R5, and R9 are all hydrogen, then R8 must be halogen; and when R4 isNOH, R6-a is methyl, R6 is hydrogen or hydroxyl, R7 is halogen, R5 andR9 are both hydrogen, then R8 must be halogen; and when R4 isN-NH-alkyl, R6-a is methyl, R6 is hydroxyl and R7, R5, R9 are allhydrogen, then R8 must be halogen; and when R4 is NH-alkyl, R6-a, R6, R5and R9 are all hydrogen, R7 is hydrogen, amino, mono(lower alkyl)amino,halogen, di(lower alkyl)amino or hydroxyl, then R8 must be halogen; andwhen R4 is NH-alkyl, R6-a is methyl, R6 and R9 are both hydrogen, R5 ishydroxyl, and R7 is mono(lower alkyl)amino or di(lower alkyl)amino, thenR8 must be halogen; and when R4 is NH-alkyl, R6-a is methyl, R6 ishydroxy or hydrogen and R7, R5, and R9 are all be hydrogen, then R8 mustbe halogen.

[0021] In yet another embodiment, the invention provides a4-dedimethylamino tetracycline compound having general formulae (I)through (IV): General Formula (I)

wherein R7, R8, and R9 taken together in each case, have the followingmeanings: R7 R8 R9 azido hydrogen hydrogen dimethylamino hydrogen azidohydrogen hydrogen amino hydrogen hydrogen azido hydrogen hydrogen nitrodimethylamino hydrogen amino acylamino hydrogen hydrogen hydrogenhydrogen acylamino amino hydrogen nitro hydrogen hydrogen(N,N-dimethyl)glycylamino amino hydrogen amino hydrogen hydrogenethoxythiocarbonylthio dimethylamino hydrogen acylamino dimethylaminohydrogen diazonium dimethylamino chloro amino hydrogen chloro aminoamino chloro amino acylamino chloro acylamino amino chloro hydrogenacylamino chloro hydrogen monoalkylamino chloro amino nitro chloro aminodimethylamino chloro acylamino dimethylamino chloro dimethylaminodimethylamino hydrogen hydrogen hydrogen hydrogen dimethylamino andGeneral Formula (II)

wherein R7, R8, and R9 taken together in each case, have the followingmeanings: R7 R8 R9 azido hydrogen hydrogen dimethylamino hydrogen azidohydrogen hydrogen amino hydrogen hydrogen azido hydrogen hydrogen nitrodimethylamino hydrogen amino acylamino hydrogen hydrogen hydrogenhydrogen acylamino amino hydrogen nitro hydrogen hydrogen(N,N-dimethyl)glycylamino amino hydrogen amino hydrogen hydrogenethoxythiocarbonylthio dimethylamino hydrogen acylamino hydrogenhydrogen diazonium hydrogen hydrogen dimethylamino diazonium hydrogenhydrogen ethoxythiocarbonylthio hydrogen hydrogen dimethylamino chloroamino amino chloro amino acylamino chloro acylamino hydrogen chloroamino amino chloro hydrogen acylamino chloro hydrogen monoalkyl aminochloro amino nitro chloro amino and General Formula (III)

wherein R8 is hydrogen or halogen and R9 is selected from the groupconsisting of nitro, (N,N-dimethyl)glycylamino, andethoxythiocarbonylthio; and General Formula (IV)

wherein R7, R8, and R9 taken together in each case, have the followingmeanings: R7 R8 R9 amino hydrogen hydrogen nitro hydrogen hydrogen azidohydrogen hydrogen dimethylamino hydrogen azido hydrogen hydrogen aminohydrogen hydrogen azido hydrogen hydrogen nitro bromo hydrogen hydrogendimethylamino hydrogen amino acylamino hydrogen hydrogen hydrogenhydrogen acylamino amino hydrogen nitro hydrogen hydrogen(N,N-dimethyl)glycylamino amino hydrogen amino diethylamino hydrogenhydrogen hydrogen hydrogen ethoxythiocarbonylthio dimethylamino hydrogenmethylamino dimethylamino hydrogen acylamino dimethylamino chloro aminoamino chloro amino acylamino chloro acylamino hydrogen chloro aminoamino chloro hydrogen acylamino chloro hydrogen monoalkylamino chloroamino nitro chloro amino and pharmaceutically acceptable andunacceptable salts thereof.

[0022] In yet another embodiment, the invention provides a tetracyclinecompound of the formulae:

[0023] wherein R7 is selected from the group consisting of hydrogen,amino, nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of hydrogen, amino,azido, nitro, acylamino, hydroxy, ethoxythiocarbonylthio, mono(loweralkyl) amino, halogen, diazonium, di(lower alkyl)amino and RCH(NH₂)CO; Ris hydrogen or lower alkyl; R^(a) and R^(b) are selected from the groupconsisting of hydrogen, methyl, ethyl, n-propyl and 1-methylethyl withthe proviso that R^(a) and R^(b) cannot both be hydrogen; R^(c) andR^(d) are, independently (CH₂)_(n)CHR^(e) wherein n is 0 or 1 and R^(e)is selected from the group consisting of hydrogen, alkyl, hydroxy,lower(C₁-C₃) alkoxy, amino, or nitro; and, W is selected from the groupconsisting of (CHR^(e))_(m) wherein m is 0-3 and R^(e) is as above, NH,N(C₁-C₃) straight chained or branched alkyl, O, S and N(C ₁-C₄) straightchain or branched alkoxy; and pharmaceutically acceptable andunacceptable salts thereof.

[0024] In a further embodiment, the following provisos apply: wheneither R7 and R9 are hydrogen then R8 must be halogen; and when R6-a,R6, R5 and R9 are all hydrogen and R7 is hydrogen, amino, nitro,halogen, dimethylamino or diethylamino, then R8 must be halogen; andwhen R6-a is methyl, R6 and R9 are both hydrogen, R5 is hydroxyl, and R7is hydrogen, amino, nitro, halogen or diethylamino, then R8 is halogen;and when R6-a is methyl, R6 is hydroxyl, R5, R7 and R9 are all hydrogen,then R8 must be halogen; and when R6-a, R6 and R5 are all hydrogen, R9is methylamino and R7 is dimethylamino, then R8 must be halogen; andwhen R6-a is methyl, R6 is hydrogen, R5 is hydroxyl, R9 is methylaminoand R7 is dimethylamino, then R8 must be halogen; and when R6-a ismethyl, R6, R5 and R9 are all hydrogen and R7 is cyano, then R8 must behalogen.

[0025] In another embodiment, the tetracycline compounds described aboveand below preferably have a PIF value from about 1 to about 2, morepreferably at about 1. Some examples of tetracycline compounds having aPIF value of about 1 include:

STRUCTURE K

[0026] wherein: R7, R8, and R9 taken together in each case, have thefollowing meanings: STRUCTURE K wherein: R7, R8, and R9 taken togetherin each case, have the following meanings: R7 R8 R9 hydrogen hydrogenamino hydrogen hydrogen palmitamide

STRUCTURE L STRUCTURE M STRUCTURE N STRUCTURE O

[0027] wherein: R7, R8, and R9 taken together in each case, have thefollowing meanings: R7 R8 R9 hydrogen hydrogen acetamido hydrogenhydrogen dimethylaminoacetamido hydrogen hydrogen nitro hydrogenhydrogen amino

STRUCTURE P

[0028] wherein: R7, R8, and R9 taken together are, respectively,hydrogen, hydrogen and nitro.

[0029] Some examples of tetracycline compounds having a PIF value fromabout 1 to about 2 have the general formula:

STRUCTURE K:

[0030] wherein: R7, R8, and R9 taken together are, respectively,hydrogen, hydrogen and dimethylamino.

[0031] In another embodiment, the present invention provides atetracycline compound of the formulae:

STRUCTURE C STRUCTURE D STRUCTURE E STRUCTURE F

[0032] wherein R7 is selected from the group consisting of aryl, alkenyland alkynyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of hydrogen, amino,azido, nitro, acylamino, hydroxy, ethoxythiocarbonylthio, mono(loweralkyl) amino, halogen, diazonium, di(lower alkyl)amino and RCH(NH₂)CO;and pharmaceutically acceptable and unacceptable salts thereof; or

STRUCTURE C STRUCTURE D STRUCTURE E STRUCTURE F

[0033] wherein: R7 is selected from the group consisting of hydrogen,amino, nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of an aryl, alkenyland alkynyl; and pharmaceutically acceptable and unacceptable saltsthereof; or

STRUCTURE C STRUCTURE D STRUCTURE E STRUCTURE F

[0034] wherein: R7 and R9 are selected from the group consisting of anaryl, alkenyl alkynyl, or mixures thereof, R6-a is selected from thegroup consisting of hydrogen and methyl; R6 and R5 are selected from thegroup consisting of hydrogen and hydroxyl; R8 is selected from the groupconsisting of hydrogen and halogen; and pharmaceutically acceptable andunacceptable salts thereof.

[0035] In another embodiment, the invention provides a tetracyclinecompound of the formulae:

STRUCTURE G STRUCTURE H STRUCTURE I STRUCTURE J

[0036] wherein R7 is selected from the group consisting of an aryl,alkenyl and alkynyl; R6-a is selected from the group consisting ofhydrogen and methyl; R6 and R5 are selected from the group consisting ofhydrogen and hydroxyl; R4 is selected from the group consisting of NOH,N-NH-A, and NH-A, where A is a lower alkyl group; R8 is selected fromthe group consisting of hydrogen and halogen; R9 is selected from thegroup consisting of hydrogen, amino, azido, nitro, acylamino, hydroxy,ethoxythiocarbonylthio, mono(lower alkyl) amino, halogen, di(loweralkyl)amino and RCH(NH₂)CO; and pharmaceutically acceptable andunacceptable salts thereof; or

STRUCTURE G STRUCTURE H STRUCTURE I STRUCTURE J

[0037] wherein R7 is selected from the group consisting of hydrogen,amino, nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R4 is selected from the group consisting of NOH, N-NH-A, andNH-A, where A is a lower alkyl group; R8 is selected from the groupconsisting of hydrogen and halogen; R9 is selected from the groupconsisting of an aryl, alkenyl and alkynyl; and pharmaceuticallyacceptable and unacceptable salts thereof, or

STRUCTURE G STRUCTURE H STRUCTURE I STRUCTURE J

[0038] wherein: R7 and R9 are selected from the group consisting of anaryl, alkenyl, alkynyl; or mixtures thereof; R6-a is selected from thegroup consisting of hydrogen and methyl; R6 and R5 are selected from thegroup consisting of hydrogen and hydroxyl; R4 is selected from the groupconsisting of NOH, N-NH-A, and NH-A, where A is a lower alkyl group; andR8 is selected from the group consisting of hydrogen and halogen; andpharmaceutically acceptable and unacceptable salts thereof;

[0039] In another embodiment, the invention provides a tetracyclinecompound of the formulae:

STRUCTURE K

[0040] wherein R7 is selected from the group consisting of an aryl,alkenyl and alkynyl; R8 is selected from the group consisting ofhydrogen and halogen; R9 is selected from the group consisting ofhydrogen, amino, azido, nitro, acylamino, hydroxy,ethoxythiocarbonylthio, mono(lower alkyl) amino, halogen, di(loweralkyl)amino and RCH(NH₂)CO; and pharmaceutically acceptable andunacceptable salts thereof; or

STRUCTURE K

[0041] wherein: R7 is selected from the group consisting of hydrogen,amino, nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of an aryl, alkenyland alkynyl; and pharmaceutically acceptable and unacceptable saltsthereof; or

STRUCTURE K

[0042] wherein: R7 and R9 are selected from the group consisting of anaryl, alkenyl, alkynyl and mixtures thereof; and R8 is selected from thegroup consisting of hydrogen and halogen; and pharmaceuticallyacceptable and unacceptable salts thereof; and

STRUCTURE L STRUCTURE M STRUCTURE N STRUCTURE O

[0043] wherein: R7 is selected from the group consisting of an aryl,alkenyl and alkynyl; R8 is selected from the group consisting ofhydrogen and halogen; and pharmaceutically acceptable and unacceptablesalts thereof; or

STRUCTURE L STRUCTURE M STRUCTURE N STRUCTURE O

[0044] wherein R7 is selected from the group consisting of hydrogen,amino, nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of an aryl, alkenyland alkynyl; and pharmaceutically acceptable and unacceptable saltsthereof; or

STRUCTURE L STRUCTURE M STRUCTURE N STRUCTURE O

[0045] wherein R7 is and R9 are selected from the group consisting of anaryl, alkenyl, alkynyl and mixtures thereof; R8 is selected from thegroup consisting of hydrogen and halogen; R9 is selected from the groupconsisting of hydrogen, amino, azido, nitro, acylamino, hydroxy,ethoxythiocarbonylthio, mono(lower alkyl) amino, halogen, di(loweralkyl)amino and RCH(NH₂)CO; and pharmaceutically acceptable andunacceptable salts thereof; and

STRUCTURE P

[0046] wherein R9 is selected from the group consisting of an aryl,alkenyl and alkynyl; and R8 is selected from the group consisting ofhydrogen and halogen; and pharmaceutically acceptable and unacceptablesalts thereof; and

STRUCTURE Q STRUCTURE R

[0047] wherein R7 is selected from the group consisting of an aryl,alkenyl and alkynyl; R8 is selected from the group consisting ofhydrogen and halogen; R9 is selected from the group consisting ofhydrogen, amino, azido, nitro, acylamino, hydroxy,ethoxythiocarbonylthio, mono(lower alkyl) amino, halogen, di(loweralkyl)amino and RCH(NH₂)CO; and pharmaceutically acceptable andunacceptable salts thereof; or

STRUCTURE Q STRUCTURE R

[0048] wherein R7 is selected from the group consisting of hydrogen,amino, nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of an aryl, alkenyland alkynyl; and pharmaceutically acceptable and unacceptable saltsthereof; or

STRUCTURE Q STRUCTURE R

[0049] wherein R7 and R9 are selected from the group consisting of anaryl, alkenyl, alkynyl; and mixtures thereof; R8 is selected from thegroup consisting of hydrogen and halogen; and pharmaceuticallyacceptable and unacceptable salts thereof.

[0050] In another embodiment, the invention provides a tetracyclinecompound of the formulae:

STRUCTURES S-Z

[0051] wherein R7 is selected from the group consisting of an aryl,alkenyl and alkynyl; R6-a is selected from the group consisting ofhydrogen and methyl; R6 and R5 are selected from the group consisting ofhydrogen and hydroxyl; R8 is selected from the group consisting ofhydrogen and halogen; R9 is selected from the group consisting ofhydrogen, amino, azido, nitro, acylamino, hydroxy,ethoxythiocarbonylthio, mono(lower alkyl) amino, halogen, diazonium,di(lower alkyl)amino and RCH(NH₂)CO; R^(a) and R^(b) are selected fromthe group consisting of hydrogen, methyl, ethyl, n-propyl and1-methylethyl with the proviso that R^(a) and R^(b) cannot both behydrogen; R^(c) and R^(d) are, independently, (CH₂)_(n)CHR^(e) wherein nis 0 or 1 and R^(e) is selected from the group consisting of hydrogen,alkyl, hydroxy, lower(C₁-C₃) alkoxy, amino, or nitro; and, W is selectedfrom the group consisting of (CHR^(e))_(m) wherein m is 0-3 and saidR^(e) is as above, NH, N(C₁-C₃) straight chained or branched alkyl, O, Sand N(C₁-C₄) straight chain or branched alkoxy; and pharmaceuticallyacceptable and unacceptable salts thereof; or

STRUCTURES S-Z

[0052] wherein R7 is selected from the group consisting of hydrogen,amino, nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of an aryl, alkenyland alkynyl; R^(a) and R^(b) are selected from the group consisting ofhydrogen, methyl, ethyl, n-propyl and 1-methylethyl with the provisothat R^(a) and R^(b) cannot both be hydrogen; R^(c) and R^(d) are,independently, (CH₂)_(n)CHR^(e) wherein n is 0 or 1 and R^(e) isselected from the group consisting of hydrogen, alkyl, hydroxy,lower(C₁-C₃) alkoxy, amino, or nitro; and, W is selected from the groupconsisting of (CHR^(e))_(m) wherein m is 0-3 and said R^(e) is as above,NH, N(C₁-C₃) straight chained or branched alkyl, O, S and N(C₁-C₄)straight chain or branched alkoxy; and pharmaceutically acceptable andunacceptable salts thereof; or

STRUCTURES S-Z

[0053] wherein: R7 and R9 are selected from the group consisting of anaryl, alkenyl, alkynyl and mixtures thereof; R6-a is selected from thegroup consisting of hydrogen and methyl; R6 and R5 are selected from thegroup consisting of hydrogen and hydroxyl; R8 is selected from the groupconsisting of hydrogen and halogen; R^(a) and R^(b) are selected fromthe group consisting of hydrogen, methyl, ethyl, n-propyl and1-methylethyl with the proviso that R^(a) and R^(b) cannot both behydrogen; R^(c) and R^(d) are, independently, (CH₂)_(n)CHR^(e) wherein nis 0 or 1 and R^(e) is selected from the group consisting of hydrogen,alkyl, hydroxy, lower(C₁-C₃) alkoxy, amino, or nitro; and W is selectedfrom the group consisting of (CHR^(e))_(m) wherein m is 0-3 and saidR^(e) is as above, NH, N(C₁-C₃) straight chained or branched alkyl, O, Sand N(C₁-C₄) straight chain or branched alkoxy; and pharmaceuticallyacceptable and unacceptable salts thereof.

[0054] The present invention includes a method for treating a mammalsuffering from a condition that benefits from a non-antimicrobial doseof a tetracycline compound. Some examples of such conditions includethose characterized by excessive collagen destruction, excessive MMPenzyme activity, excessive TNF activity, excessive nitric oxideactivity, excessive IL-1 activity, excessive elastase activity,excessive loss of bone density, excessive protein degradation, excessivemuscle wasting, excessive glycosylation of collagen, excessive COX-2activity, insufficient bone protein synthesis, insufficientinterleukin-10 production, or excessive phospholipase A₂ activity. Themethod for treating comprises administering to the mammal an effectiveamount of a tetracycline compound of the invention.

[0055] Conditions that benefit from a non-antimicrobial dose of atetracycline compound include, but are not limited to, abdominal aorticaneurysm, ulceration of the cornea, periodontal disease, diabetes,diabetes mellitus, scleroderma, progeria, lung disease, cancer, graftversus host disease, disease of depressed bone marrow function,thrombocytopenia, prosthetic joint loosening, spondyloarthropathies,osteoporosis, Paget's disease, autoimmune disease, systemic lupuserythematosus, acute or chronic inflammatory condition, renal disease,connective tissue disease or neurological and neurodegenerativeconditions.

[0056] Acute or chronic inflammatory conditions that can benefit from anon-antimicrobial dose of a tetracycline compound can be, but are notlimited to, inflammatory bowel disease, arthritis, osteoarthritis,rheumatoid arthritis, pancreatitis, nephritis, glomerulonephritis,sepsis, septic shock, lipopolysaccharide endotoxin shock, multisystemorgan failure or psoriasis.

[0057] The lung diseases that can benefit from a non-antimicrobial doseof a tetracycline compound can be, but are not limited to, ARDS, cysticfibrosis, emphysema or acute lung injury resulting from inhalation oftoxicants.

[0058] The renal diseases that can benefit from a non-antimicrobial doseof a tetracycline compound include, but are not limited to chronic renalfailure, acute renal failure, nephritis or glomerulonephritis.

[0059] These and other advantages of the present invention will beappreciated from the detailed description and examples which are setforth herein. The detailed description and examples enhance theunderstanding of the invention, but are not intended to limit the scopeof the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

[0060]FIG. 1 shows the photoirritancy factor (PIF), also known as thephotoinhibition factor, for some tetracycline compounds. For structureK, the compounds indicated are as follows: COL R7 R8 R9 308 hydrogenhydrogen amino 311 hydrogen hydrogen palmitamide 306 hydrogen hydrogendimethylamino

[0061] For structures L, M, N or O the compounds indicated are asfollows: COL R7 R8 R9 801 hydrogen hydrogen acetamido 802 hydrogenhydrogen dimethylaminoacetamido 804 hydrogen hydrogen nitro 805 hydrogenhydrogen amino

[0062] For structure P, R7 is hydrogen, R8 is hydrogen and R9 is nitro.

DETAILED DESCRIPTION OF THE INVENTION

[0063] Particularly preferred compounds of the present invention have Dring substituents at the 7 and/or 9 positions on the4-dedimethylaminotetracycline molecule. These compounds include7-azido-6-demethyl-6-deoxy-4-dedimethylamino tetracycline,7-dimethylamino-9-azido-6-demethyl-6-deoxy-4-dedimethylaminotetracycline, 9-amino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-azido-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-nitro-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-amino-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,7-acetamido-6-demethyl-6-deoxy-4-dedimethylamino tetracycline,9-acetamido-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,7-amino-9-nitro-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-(N,N,-dimethyl)glycylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,7,9-diamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-ethoxythiocarbonylthio-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,7-dimethylamino-9-acetamido-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-azido-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-azido-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,7-azido-5-hydroxy-6-deoxy-4-dedimethylamino tetracycline,7-dimethylamino-9-azido-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline, 9-amino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,9-azido-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,9-nitro-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,9-amino-7-dimethylamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,7-acetamido-5-hydroxy-6-deoxy-4-dedimethylamino tetracycline,9-acetamido-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,7-amino-9-nitro-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,9-(N,N-dimethyl)glycylamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,7,9-diamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,7-dimethylamino-9-amino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,9-ethoxythiocarbonylthio-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,7-dimethylamino-9-acetamido-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,9-azido-7-dimethylamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,9-amino-8-chloro-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,9-(N,N-dimethyl)glycylamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,9-nitro-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-acetamido-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-(N,N-dimethyl)glycylamino-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,and9-ethoxythiocarbonylthio-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline.

[0064] In addition, the D ring may be halogenated at the C8 position toprovide 8-halodedimethylaminotetracycline derivatives. As used in thisspecification, halogens can be chlorine, fluorine, bromine, and iodine.Some examples of 8-halo-dedimethylaminotetracycline derivatives are9-amino-8-chloro-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-amino-8-chloro-7-dimethylamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracyclineand 9-amino-8-chloro-6-demethyl-6-deoxy-4-dedimethylaminotetracycline.

[0065] In one embodiment of the invention, the4-dedimethylaminotetracycline derivatives are substituted with an oxime,NH-alkyl, or N-NH-alkyl group at the C4 position. These compounds havethe general formula:

STRUCTURE G STRUCTURE H STRUCTURE I STRUCTURE J

[0066] wherein R7 is selected from the group consisting of hydrogen,amino, nitro, mono(lower alkyl) amino, halogen, and di(loweralkyl)amino, ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano,and hydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R4 is selected from the group consisting of NOH, N-NH-A, andNH-A, where A is a lower alkyl group; R8 is selected from the groupconsisting of hydrogen and halogen; R9 is selected from the groupconsisting of hydrogen, amino, azido, nitro, acylamino, hydroxy,ethoxythiocarbonylthio, mono(lower alkyl) amino, halogen, di(loweralkyl)amino and RCH(NH₂)CO; R is hydrogen or lower alkyl; andpharmaceutically acceptable and unacceptable salts thereof; with thefollowing provisos: when R4 is NOH, N-NH-alkyl or NH-alkyl and R7, R6-a,R6, R5, and R9 are all hydrogen, then R8 must be halogen; and when R4 isNOH, R6-a is methyl, R6 is hydrogen or hydroxyl, R7 is halogen, R5 andR9 are both hydrogen, then R8 must be halogen; and when R4 isN-NH-alkyl, R6-a is methyl, R6 is hydroxyl and R7, R5, R9 are allhydrogen, then R8 must be halogen; and when R4 is NH-alkyl, R6-a, R6, R5and R9 are all hydrogen, R7 is hydrogen, amino, mono(lower alkyl)amino,halogen, di(lower alkyl)amino or hydroxyl, then R8 must be halogen; andwhen R4 is NH-alkyl, R6-a is methyl, R6 and R9 are both hydrogen, R5 ishydroxyl, and R7 is mono(lower alkyl)amino or di(lower alkyl)amino, thenR8 must be halogen; and when R4 is NH-alkyl, R6-a is methyl, R6 ishydroxy or hydrogen and R7, R5, and R9 are all be hydrogen, then R8 mustbe halogen.

[0067] It will be understood that if the stereochemistry of asubstituent on rings A-D of the novel 4-dedimethylaminotetracyclinederivative is not specified, then both epimers are intended to beencompassed.

[0068] As used herein, NH-Alkyl, N-NH-Alkyl, alkoxy and alkyl groupscontain straight or branched, saturated or unsaturated alkyl carbonchains, having from one to twenty-six carbon atoms. For example, alkylgroups include fatty alkyls which contain ten to twenty-six carbonatoms. Some examples of saturated fatty alkyl groups include, lauryl,myristyl, palmityl, stearyl, etc. Some examples of unsaturated fattyalkyl groups include palmitoleyl, oleyl, linoleyl, linolenyl, etc.

[0069] Alkyl groups also include lower alkyls which include straight orbranched, saturated or unsaturated carbon chains, having from one to sixcarbon atoms. Some examples of lower alkyl groups are methyl, ethyl,propyl, butyl, isobutyl, n-butyl, secondary butyl, tertiary butyl,n-pentyl and benzyl. The alkyl moiety of acyl groups is defined asabove. Some examples of acyl groups include acetyl, propionyl, butyryl,and acyl groups comprising fatty acids such as those described above.

[0070] Preferred oxime compounds include7-azido-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7-dimethylamino-9-azido-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-amino-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-azido-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-nitro-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-amino-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7-acetamido-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-acetamido-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7-amino-9-nitro-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-(N,N,-dimethyl)glycylamino-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7,9-diamino-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-ethoxythiocarbonylthio-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7-dimethylamino-9-acetamido-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-azido-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-azido-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7-azido-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7-dimethylamino-9-azido-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximino-tetracycline,9-amino-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-azido-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-nitro-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-amino-7-dimethylamino-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7-acetamido-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-acetamido-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7-amino-9-nitro-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-(N,N-dimethyl)glycylamino-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,also known as9-dimethylaminoacetamido-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7,9-diamino-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7-dimethylamino-9-amino-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-ethoxythiocarbonylthio-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,7-dimethylamino-9-acetamido-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-azido-7-dimethylamino-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-amino-8-chloro-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-(N,N-dimethyl)glycylamino-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-nitro-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-acetamido-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylamino-4-oximino-tetracycline,9-(N,N-dimethyl)glycylamino-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylamino-4-oximino-tetracycline,and9-ethoxythiocarbonylthio-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline.

[0071] In addition, the D ring may be halogenated at the C8 position toprovide 8-halo-4-dedimethylamino-4-oximinotetracycline compounds. Someexamples include9-amino-8-chloro-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline,9-amino-8-chloro-7-dimethylamino-5-hydroxy-6-deoxy-4-dedimethylamino-4-oximinotetracyclineand9-amino-8-chloro-6-demethyl-6-deoxy-4-dedimethylamino-4-oximinotetracycline.

[0072] Preferred hydrazone compounds include7-azido-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,7-dimethylamino-9-azido-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,9-amino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,9-azido-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,9-nitro-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,9-amino-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,7-acetamido-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,9-acetamido-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,7-amino-9-nitro-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,9-(N,N,-dimethyl)glycylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,7,9-diamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,9-ethoxythiocarbonylthio-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,7-dimethylamino-9-acetamido-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,9-azido-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methyl or ethyl hydrazone,9-azido-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,7-azido-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,7-dimethylamino-9-azido-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,9-amino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,9-azido-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone, 9-nitro-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methyl or ethyl hydrazone,9-amino-7-dimethylamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,7-acetamido-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,9-acetamido-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,7-amino-9-nitro-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone, 9-(N,N-dimethyl)glycylamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,7,9-diamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methyl orethyl hydrazone,7-dimethylamino-9-amino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,9-ethoxythiocarbonylthio-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,7-dimethylamino-9-acetamido-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,9-azido-7-dimethylamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,9-amino-8-chloro-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone, 9-(N,N-dimethyl)glycylamino-5-hydroxy-6-deoxy-4-dedimethylamino tetracycline-4-methyl orethyl hydrazone,9-nitro-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone,9-acetamido-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone, 9-(N,N-dimethyl)glycylamino-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone, and9-ethoxythiocarbonylthio-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone.

[0073] The D ring may be halogenated at the C8 position to provide8-halo-4-dedimethylaminotetracycline-4-hydrazone compounds. Someexamples include9-amino-8-chloro-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methyl or ethyl hydrazone,9-amino-8-chloro-7-dimethylamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline-4-methyl or ethyl hydrazone and9-amino-8-chloro-6-demethyl-6-deoxy-4-dedimethylaminotetracycline-4-methylor ethyl hydrazone.

[0074] Novel 4-dedimethlyaminotetracycline derivatives of the presentinvention also include compounds with an NH-Alkyl (alkylamino)substituent at the C4 position on the A ring. These compounds havesubstitutions at the C5, C6, C6a, C7, C8 and/or C9 positions asdescribed above. An example is9-azido-8-chloro-7-dimethylamino-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline.

[0075] In addition, a hexanoylamino group can be added to the C9position on the D ring of any compound of the invention. An exampleincludes, but is not limited to,4-dedimethylamino-6-demethyl-6-deoxy-9-hexanoylaminotetracycline.

[0076] In another embodiment of the invention, the4-dedimethylaminotetracycline derivatives are Mannich derivatives of thecompounds described above. Such derivatives include, for example,compounds having the general formula:

STRUCTURE S STRUCTURE T STRUCTURE U STRUCTURE V

STRUCTURE W STRUCTURE X STRUCTURE Y STRUCTURE Z

[0077] wherein R5, R6, R7, R8 and R9 are as described above; R^(a) andR^(b) are selected from the group consisting of hydrogen, methyl, ethyl,n-propyl or 1-methylethyl with the proviso that R^(a) and R^(b) cannotboth be hydrogen; R^(c) and R^(d) are, independently, (CH₂)_(n)CHR^(e)wherein n is 0 or 1 and R^(e) is selected from the group consisting ofhydrogen, alkyl, hydroxy, lower(C₁-C₃) alkoxy, amino, or nitro; and, Wis selected from the group consisting of (CHR^(e))_(m) wherein m is 0-3and R^(e) is as above, NH, N(C₁-C₃) straight chained or branched alkyl,O, S and N(C₁-C₄) straight chain or branched alkoxy; andpharmaceutically acceptable and unacceptable salts thereof. For example,when m is 0, R^(c) and R^(d) are bonded to each other in a 3-5 memberedring, such as, for example, a pyrrolidino or substituted pyrrolidinoring, a morpholino or substituted morpholino ring, or a piperazino orsubstituted piperazino ring.

[0078] These Mannich derivatives include, for example, compounds with apiperazin-1-yl, 4-methylpiperazin-1-yl, morpholin-1-yl, orpyrrolidin-1-yl substituent at the C2 position. These compounds havesubstituents at the C4, C5, C6, C6a, C7, C8 and/or C9 positions asdescribed above. Examples of such compounds include, but are not limitedto,N-morpholin-1-ylmethyl-4-dedimethylamino-6-demethyl-6-deoxytetracyline,N-pyrrolidin-1-ylmethyl-4-dedimethylamino-6-demethyl-6-deoxytetracyline,N-morpholin-1-ylmethyl-4-dedimethylamino-6-demethyl-6-deoxy-9-hexanoylaminotertracycline,N-pyrrolidin-1-ylmethyl-4-dedimethylamino-6-demethyl-6-deoxy-9-hexanoylaminotetracycline.

[0079] In another embodiment of the invention, the4-dedimethylaminotetracycline derivatives are substituted at the C7 orC9 position, or at both the C7 and C9 positions, with an aryl, alkenyl,or alkynyl group, or mixtures thereof. Such compounds include, forexample, compounds having any of the general formulas C-Z wherein R4,R5, R6, R6a, R8, Ra, Rb, Rc, Rd and W are as described above; with theprovisos that when R7 is not an aryl, alkenyl or alkynyl group, R7 is ahydrogen, amino, nitro, mono(lower alkyl) amino, halogen, di(loweralkyl)amino, ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano,or hydroxyl; and when R9 is not an aryl, alkenyl or alkynyl group, R9 ishydrogen, amino, azido, nitro, acylamino, hydroxy,ethoxythiocarbonylthio, mono(lower alkyl)amino, halogen, di(loweralkyl)amino or RCH(NH₂)CO.

[0080] In this specification, an aryl group is any monocyclic orpolycyclic aromatic group derived from an aromatic compound. The mosttypical example of a monocyclic aromatic compound is benzene andsubstituted benzene derivatives. Examples of polycyclic aromaticcompounds include, but are not limited to, naphthalene, anthracene,1,2-benzylpyrene, and coronene.

[0081] The alkenyl, and alkynyl groups at either or both of the C7 or C9positions include any of the alkyl groups described herein, furtherhaving, respectively, one or more double or triple bonds, preferably oneto three double or triple bonds, at any position. Some examples ofalkenyl groups include, but are not limited to, ethylenyl, propenyl,1-butylenyl, 2-butylenyl and 2-methylpropylenyl and 1,3 hexadienyl. Thealkynyl groups include, but are not limited to, ethynyl, propynyl,1-butynyl, 2-butynyl and 2-methylpropynyl and 1,3 hexadiynyl.

[0082] The aryl, alkenyl and alkynyl groups may be further substitutedat any position with one or more additional substituents. Some examplesof such further substitutions include, but are not limited to, nitro,amino, halo (F, Cl, Br or I), amido, azido, cyano, hydroxyl, alkoxy,preferably lower alkoxy, acyl, preferably lower acyl, amidoazido,mono(lower alkyl)amino, di(lower alkyl)amino, ethoxythiocarboxylthio,diazonium, acylamino, N,N-dimethylglycylamino, and alkyl groups,preferably lower alkyl groups. Examples of substituted benzenederivatives include, but are not limited to, methylbenzene (toluene),nitrobenzene, hydroxybenzene (phenol), aminobenzene (aniline),vinylbenzene (styrene), benzaldehyde, benzoic acid, 1,2-dimethylbenzene(ortho-xylene), 1,3 dimethylbenzene (meta-xylene), and1,4-dimethylbenzene (para-xylene).

[0083] Some examples of compounds having aryl substituents at the C7 orC9 positions include7-phenyl-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-phenyl-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,7-(4-fluorophenyl)-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-(4-fluorphenyl)-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,7-(4-chlorophenyl)-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-(4-chlorophenyl)-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,7-(4-nitrophenyl)-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-(4-nitrophenyl)-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,7-(4-dimethylamino)-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,9-(4-dimethylamino)-6-demethyl-6-deoxy-4-dedimethylaminotetracycline,7-phenyl-5-hydroxy-6-deoxy-4-dedimethyaminotetracycline,9-phenyl-5-hydroxy-6-deoxy-4-dedimethylaminotetracycline,7-(4-fluorophenyl)-5-hydroxy-6-deoxy-4-dedimethyaminotetracycline,9-(4-fluorphenyl)-5-hydroxy-6-deoxy-4-dedimethyaminotetracycline,7-(4-chlorophenyl)-5-hydroxy-6-deoxy-4-dedimethyaminotetracycline,9-(4-chlorophenyl)-5-hydroxy-6-deoxy-4-dedimethyaminotetracycline,7-(4-nitrophenyl)-5-hydroxy-6-deoxy-4-dedimethyaminotetracycline,9-(4-nitrophenyl)-5-hydroxy-6-deoxy-4-dedimethyaminotetracycline,7-(4-dimethylamino)-5-hydroxy-6-deoxy-4-dedimethyaminotetracycline,9-(4-dimethylamino)-5-hydroxy-6-deoxy-4-dedimethyaminotetracycline,7-phenyl-7-dimethylamino-4-dedimethylaminotetracycline,9-phenyl-7-dimethylamino-4-dedimethylaminotetracycline,7-(4-fluorophenyl)-7-dimethylamino, 4-dedimethylaminotetracycline,9-(4-fluorphenyl)-7-dimethylamino-4-dedimethylaminotetracycline,7-(4-chlorophenyl)-7-dimethylamino-4-dedimethylaminotetracycline,9-(4-chlorophenyl)-7-dimethylamino-4-dedimethylaminotetracycline,7-(4-nitrophenyl)-7-dimethylamino-4-dedimethylaminotetracycline,9-(4-nitrophenyl)-7-dimethylamino-4-dedimethylaminotetracycline,7-(4-dimethylamino)-7-dimethylamino-4-dedimethylaminotetracycline,9-(4-dimethylamino)-7-dimethylamino-4-dedimethylaminotetracycline,7-phenyl-4-dedimethylaminotetracycline,9-phenyl-4-dedimethylaminotetracycline,7-(4-fluorophenyl)-4-dedimethylaminotetracycline,9-(4-fluorphenyl)-4-dedimethylaminotetracycline,7-(4-chlorophenyl)-4-dedimethylaminotetracycline,9-(4-chlorophenyl)-4-dedimethylaminotetracycline,7-(4-nitrophenyl)-4-dedimethylaminotetracycline,9-(4-nitrophenyl)-4-dedimethylaminotetracycline,7-(4-dimethylamino)-4-dedimethylaminotetracycline,9-(4-dimethylamino)-4-dedimethylaminotetracycline.

[0084] Some examples of compounds having alkenyl or alkynyl substituentsat the C7 or C9 positions include the same C7 and C9 aryl substitutedtetracycline derivatives described above, but instead having alkenyl oralkynyl substituents that are two to four carbons in length attached tothe C7 and C9 positions. The alkenyl groups include, but are not limitedto, ethenyl, propenyl, 1-butenyl, 2-butenyl and 2-methylpropenyl. Thealkynyl groups include, but are not limited to, ethynyl, propynyl,1-butynyl, 2-butynyl and 2-methylpropynyl.

[0085] The novel 4-dedimethylaminotetracycline compounds of the presentinvention including pharmaceutically acceptable and unacceptable saltsthereof may be prepared by D ring substitution at the C7, C8 and/or C9positions using starting reactants that can readily be prepared orpurchased by methods known in the art. See, for example, Mitscher, L.A., The Chemistry of the Tetracycline Antibiotics, Marcel Dekker, NewYork (1978), Ch. 6, Hlavka, J. and J. H. Boothe, The Tetracyclines,Springer-Verlag, Berlin-Heidelberg, page 18 (1985) and U.S. Pat. Nos.4,704,383, 3,226,436, 3,047,626 3,518,306 and 5,532,227.

[0086] For example, nitration of the C9 position on the D ring may beaccomplished, and novel 9-nitro compounds may be prepared, by usingknown starting reactants such as7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline or6-deoxy-4-dedimethylaminotetracycline and treating these compounds witha strong acid and metal nitrate salts. Examples of strong acids that aresuitable for use in the present invention are: sulfuric acid,trifluoroacetic acid, methanesulfonic acid or perchloric acid. Suitablemetal nitrate salts are, for example, calcium, potassium or sodiumnitrate. The C9 position on the D ring undergoes nitration to form thecorresponding9-nitro-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracyclineor 9-nitro-6-deoxy-4-dedimethylaminotetracycline compounds.

[0087] Amination of the C9 position on the D ring may be accomplished bytreating a 9-nitro-4-dedimethylaminotetracycline, such as9-nitro-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracyclineor 9-nitro-6-deoxy-4-dedimethylaminotetracycline with hydrogen in thepresence of a suitable supported catalyst such as Raney nickel, platinumoxide or palladium-on-carbon. This is then filtered and washed with anorganic solvent such as ether. The C9 substituent is reduced to form thecorresponding9-amino-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracyclineor 9-amino-6-deoxy-4-dedimethylaminotetracycline compound.

[0088] The amino group on the D ring at the C9 position may be convertedto an acylamido group or a sulfonamido group. For example,9-amino-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracyclineor 9-amino-6-deoxy-4-dedimethylaminotetracycline compounds are treatedwith acyl chloride, acyl anhydride, mixed acyl anhydride, sulfonylchloride or sulfonyl anhydride in the presence of a suitable acidscavenger dispersed in a solvent. The acid scavenger is suitablyselected from sodium bicarbonate, sodium acetate, pyridine,triethylamine, N,O-bis(trimethylsilyl)acetamide,N,O-bis(trimethylsilyl)trifluoroacetamide or a basic ion-exchange resin.Solvents suitable for the acylation reaction include water,water-tetrahydrofuran, N-methylpyrolidone,1,3-dimethyl-2-imidazolidione, hexamethylphosphoramide,1,3,dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone or1,2-dimethoxyethane. The C9 amino group may be converted to theacetamido group to form, for example, the corresponding9-acetamido-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline or 9-acetamido-6-deoxy-4-dedimethylamino tetracycline.

[0089] A diazonium group can also be substituted at the C9 position onthe D-ring. Typically, a 9-amino-4-dedimethylaminotetracyclinederivative, such as9-amino-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracyclineor 9-amino-6-deoxy-4-dedimethylaminotetracycline in 0. 1N HCL inmethanol is treated with n-butyl nitrite to form the corresponding9-diazonium derivatives such as9-diazonium-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracyclineor 9-diazonium-6-deoxy-4-dedimethylaminotetracycline.

[0090] The 9-diazonium-4-dedimethylaminotetracycline derivatives, suchas9-diazonium-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracyclineor 9-diazonium-6-deoxy-4-dedimethylaminotetracycline can be treated withmethanolic hydrochloric acid plus a triazo compound such as sodium azideto form, 9-azido derivatives, such as9-azido-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracyclineor 9-azido-6-deoxy-4-dedimethylaminotetracycline.

[0091] Alternately, an ethoxythiocarbonylthio group can be substitutedat the C9 position on the D ring. For example, a9-diazonium-4-dedimethylaminotetracycline derivative, such as9-diazonium-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline or 9-diazonium-6-deoxy-4-dedimethylaminotetracycline istreated with an acid metal salt such as potassium ethyl xanthate to formthe corresponding 9-ethoxythiocarbonylthio derivative, such as9-ethoxythiocarbonylthio-7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminoor 9-ethoxythiocarbonylthio-6-deoxy-4-dedimethylaminotetracycline.

[0092] The above reactions describe substitution at the C9 position on4-dedimethylaminotetracycline molecule. Some substitution may alsooccur, depending on the starting reactants and conditions used, at theC7 position and lead to also 7-substituted-4-dedimethylaminotetracyclinederivatives, such as7-diazonium-6-demethyl-6-deoxy-4-dedimethylaminotetracycline or7-azido-6-demethyl-6-deoxy-4-dedimethylaminotetracycline. The7-substituted derivatives can be separated from the 9-substitutedderivatives, and purified as discussed below.

[0093] The novel 7 or 9 azido-4-dedimethylamino derivatives of thepresent invention can be halogenated at the C8 position by treating 7 or9-azido-4-dedimethylaminotetracycline with a strong acid such ashydrogen halide, sulfuric acid saturated with hydrogen halide ormethanesulfonic acid saturated with hydrogen halide. The product that isisolated, when the hydrogen halide is hydrogen chloride, is the 8-chloro(7 or 9) amino-4-dedimethylaminotetracycline derivative. A particularlypreferred halogenated compound is9-acetamido-8-chloro-7-dimethylamino-6-deoxy-6-demethyl-4-dedimethylaminotetracycline.

[0094] In one embodiment, 4-dedimethylaminotetracycline compounds of thepresent invention have an oxime (NOH), alkylamino (NH-alkyl), oralkylhydrazone (N-NH-alkyl) group at the C4 position on the A ring.These compounds can be made using known methods. For example,4-hydroxytetracycloxide may be treated with hydroxyamine orethylhydrazine under alkaline conditions in a solvent such as methanolor ethanol. Substitution at the C4 position occurs and4-dedimethylamino-4-oximinotetracycline and4-dedimethylaminotetracycline-4-alkylhydrazone compounds can be isolatedas alkali metal salts. See for example, U.S. Pat. Nos. 3,622,627,3,159,675 and 3,345,370. Substitution at C7, C8, and/or C9 positions onthe D ring using methods previously described (i.e. halogenation,amination, or nitration) give rise to the novel 4-oxime, 4-hydrazone and4-aminoalkyl compounds of the present invention.

[0095] The Mannich derivatives can be made by methods known in the art.For example, the tetracycline derivatives described above may be treatedwith formaldehyde and the appropriate amine.

[0096] Aryl, alkenyl and alkynyl groups can be added onto the C7 and C9positions of the tetracycline derivatives described above by methodsknown in the art. Such methods include modified Suzuki and Stillecoupling reactions. See, for example, Koza D. J., Organic Letters, 2000.For example, the tetracycline derivatives described above having aniodine attached to the C7or C9 positions can be treated withtri-n-butylstannyl reagent in the presence of a palladium catalyst andcopper iodide. This reaction can be extended to a variety of aryl,alkenyl and alkynyl derivatives.

[0097] Examples of specific embodiments are described above asderivatives of the specific antibiotic compound tetracycline. Thecompounds of the invention are not, however, limited to derivatives ofany specific tetracycline compound. Rather, the compounds of theinvention include the 4-dedimethyl derivatives of any member of thetetracycline family. Thus, the invention also includes, but is notlimited to, the same 4-dedimethylamino derivatives and 4-substituted4-dedimethylamino derivatives of sancycline, minocycline, anddoxycycline as the tetracycline derivatives mentioned above.

[0098] The present invention embraces salts, including acid-addition andmetal salts, of the 4-dedimethylaminotetracycline compounds describedherein. Such salts are formed by well known procedures with bothpharmaceutically acceptable and pharmaceutically unacceptable acids andmetals. By “pharmaceutically acceptable” is meant those salt-formingacids and metals which do not substantially increase the toxicity of thecompound.

[0099] Some examples of suitable salts include salts of mineral acidssuch as hydrochloric, hydriodic, hydrobromic, phosphoric,metaphosphoric, nitric and sulfuric acids, as well as salts of organicacids such as tartaric, acetic, citric, malic, benzoic, glycollic,gluconic, gulonic, succinic, arylsulfonic, e.g. p-toluenesulfonic acids,and the like. The pharmaceutically unacceptable acid addition salts,while not useful for therapy, are valuable for isolation andpurification of the new substances. Further, they are useful for thepreparation of pharmaceutically acceptable salts. Of this group, themore common salts include those formed with hydrofluoric and perchloricacids. Hydrofluoride salts are particularly useful for the preparationof the pharmaceutically acceptable salts, e.g. the hydrochlorides, bydissolution in hydrochloric acid and crystallization of thehydrochloride salt formed. The perchloric acid salts are useful forpurification and crystallization of the new products.

[0100] Whereas metal salts may, in general, be prepared and are usefulfor various purposes, the pharmaceutically acceptable metal salts areparticularly valuable because of their utility in therapy. Thepharmaceutically acceptable metals include more commonly sodium,potassium and alkaline earth metals of atomic number up to and including20, i.e., magnesium and calcium and additionally, aluminum, zinc, ironand manganese, among others. Of course, the metal salts include complexsalts, i.e. metal chelates, which are well recognized in thetetracycline art.

[0101] After preparation, the novel compounds of the present inventioncan be conveniently purified by standard methods known in the art. Somesuitable examples include crystallization from a suitable solvent orpartition-column chromatography.

[0102] The novel 4-dedimethylaminotetracycline compounds of the presentinvention can be used in vivo, in vitro, and ex vivo, for example, inliving mammals as well as in cultured tissue, organ or cellular systems.Mammals include, for example, humans, as well as pet animals such asdogs and cats, laboratory animals, such as rats and mice, and farmanimals, such as horses and cows. Tissues, as used herein, are anaggregation of similarly specialized cells which together performcertain special functions. Cultured cellular systems include anymammalian cells, such as epithelial, endothelial, red blood, and whiteblood cells. More particularly, human peripheral blood monocytes,synovial fibroblastoid cells, and the like.

[0103] The present invention is directed to a method for treating amammal suffering from a condition or diseases that benefits from anon-antimicrobial dose of a tetracycline compound. These conditions ordiseases are characterized by excessive collagen destruction, excessiveMMP enzyme activity, excessive TNF activity, excessive nitric oxideactivity, excessive IL-1 activity, excessive elastase activity,excessive loss of bone density, excessive protein degradation, excessivemuscle wasting, excessive glycosylation of collagen, excessive COX-2activity, insufficient bone protein synthesis, insufficient IL-10(interleukin-10) production or excessive phospholipase A₂ activity. Themethod comprises administering to the mammal an effective amount of atetracycline compound of the invention.

[0104] The term “excessive,” as used herein, refers to increasedactivity over usual activity which leads to some pathological problem ina mammal or mammalian cells.

[0105] In vivo practice of the invention permits application in therelief or palliation of medical and veterinary diseases, conditions, andsyndromes. In particular, the present invention includes a method fortreating a mammal suffering from conditions or diseases includingabdominal aortic aneurysm, ulceration of the cornea, periodontaldisease, diabetes, diabetes mellitus, scleroderma, progeria, lungdisease, cancer, graft versus host diseases, disease of depressed bonemarrow function, thrombocytopenia, prosthetic joint loosening,spondyloarthropathies, osteoporosis, Paget's disease, autoimmunedisease, systemic lupus erythematosus, acute or chronic inflammatoryconditions, renal disease or connective tissue disease by administeringan effective amount of a tetracycline compound to the mammal.

[0106] Cancerous conditions treatable by tetracycline compounds of thepresent invention include, but are not limited to, carcinomas,blastomas, sarcomas such as Kaposi's Sarcoma, gliomas, and the twelvemajor cancers: prostrate cancer, breast cancer, lung cancer, colorectalcancer, bladder cancer, non-Hodgkin's lymphoma, uterine cancer,melanoma, kidney cancer, leukemia, ovarian cancer and pancreatic cancer.

[0107] Acute or chronic inflammatory conditions treatable bytetracycline compounds of the present invention include, for example,inflammatory bowel disease, arthritis, osteoarthritis, rheumatoidarthritis, pancreatitis, nephritis, glomerulonephritis, sepsis, septicshock, lipopolysaccharide endotoxin shock, multisystem organ failure orpsoriasis.

[0108] Lung diseases treatable by means of the present inventioninclude, for example, ARDS (adult respiratory distress syndrome), cysticfibrosis, emphysema or acute lung injury resulting from inhalation oftoxicants. Some examples of toxicants are acids, chemicals, industrialand military poisons, smoke and other toxic products of combustion.

[0109] The novel tetracycline compounds of the present invention canalso be used to treat renal diseases. Some examples of renal diseasesare chronic renal failure, acute renal failure, nephritis orglomerulonephritis.

[0110] The tetracycline compounds of the present invention can also beused to treat neurological and neurodegenerative conditions. Examples ofneurological and neurodegenerative conditions include, Alzheimer'sdisease, Guillain-Barré Syndrome (acute febrile polyneuritis), Krabbe'sdisease (leukodystrophy), adrenoleukodystrophy, Parkinson's disease,Huntington's disease, multiple sclerosis, amyotrophic lateral sclerosis(Lou Gehrig's disease) and encephalopathies, including spongiformencephalopathy, such as Creutzfeldt-Jakob Disease. Other examples ofspongiform encephalopathies in humans include kuru,Gerstmann-Straussler-Scheinker disease and fatal familial insomnia.Other examples of spongiform encephalopathies in mammals include bovinespongiform encephalopathy (mad cow disease), scrapie in sheep and goats,transmissible mink encephalopathy, chronic wasting disease of mule deerand elk, and feline spongiform encephalopathy.

[0111] An effective amount of a tetracycline compound as used herein isthat amount effective to achieve the specified result of treating thedisease or condition. Preferably, the tetracycline compound orderivative is provided in an amount which has little or no antimicrobialactivity. A tetracycline compound or derivative is not effectivelyantimicrobial if it does not significantly prevent the growth ofmicrobes. Accordingly, the method can beneficially employ a tetracyclinederivative which has been modified chemically to reduce or eliminate itsantimicrobial properties. The use of such chemically-modifiedtetracyclines is preferred in the present invention since they can beused at higher levels than antimicrobial tetracyclines, while avoidingcertain disadvantages, such as the indiscriminate killing of beneficialmicrobes, and the emergence of resistant microbes, which oftenaccompanies the use of antimicrobial or antibacterial amounts of suchcompounds.

[0112] The maximal dosage for a mammal is the highest dosage which doesnot cause undesirable or intolerable side effects. Minimal dosage is thelowest dosage where efficacy is first observed. For example, thetetracycline compound can be administered in an amount of from about 0.1mg/kg/day to about 30 mg/kg/day, and preferably from about 1 mg/kg/dayto about 18 mg/kg/day. In any event, the practitioner is guided by skilland knowledge in the field, and the present invention includes withoutlimitation dosages which are effective to achieve the described effect.

[0113] The method involves administering or providing a tetracyclinederivative in an amount which is effective for treating diseases orconditions in mammalian cells or a mammal. Administering thetetracycline derivatives can be accomplished in a variety of ways. Incultured cellular systems (in vitro), tetracycline derivatives can beadministered by contacting the cells directly with an effective amountof the tetracycline derivative.

[0114] In living mammals (in vivo), tetracycline derivatives of thepresent invention can be administered systemically by the parenteral andenteral routes which also includes controlled release delivery systems.For example, tetracycline derivatives of the present invention caneasily be administered intravenously (e.g., intravenous injection) whichis a preferred route of delivery. Intravenous administration can beaccomplished by mixing the tetracycline derivatives in a suitablepharmaceutical carrier (vehicle) or excipient as understood bypractitioners in the art.

[0115] Oral or enteral use is also contemplated, and formulations suchas tablets, capsules, pills, troches, elixirs, suspensions, syrups,wafers, chewing gum and the like can be employed to provide thetetracycline derivative.

[0116] Alternatively, delivery of the tetracycline derivative caninclude topical application. Accordingly, the carrier is preferablysuited for topical use. Compositions deemed to be suited for suchtopical use include gels, salves, lotions, creams ointments and thelike. The tetracycline derivative may also be incorporated with asupport base or matrix or the like to provide a pre-packaged surgical orburn dressing or bandage which can be directly applied to skin. Topicalapplication of tetracycline derivatives in amounts of up to about 25%(w/w) in a vehicle are therefore appropriate depending upon indication.More preferably, application of tetracycline derivatives in amounts offrom about 0.1% to about 10% is believed to be effective in treatingdiseases or conditions. It is believed that these quantities do notinduce significant toxicity in the subject being treated.

[0117] For example, in certain cases tetracycline compounds having onlylimited biodistribution may be preferred for localized activity. Topicalapplication of these non-absorbable CMTs would be desirable in orallesions, since the CMTs would not be absorbed to any significant degreeeven if swallowed.

[0118] Combined or coordinated topical and systemic administration oftetracycline derivatives is also contemplated under the invention. Forexample, a non-absorbable tetracycline compound can be administeredtopically, while a tetracycline compound capable of substantialabsorption and effective systemic distribution in a subject can beadministered systemically.

[0119] Photoxicity

[0120] In one embodiment, the invention relates to a class of compoundsthat have low phototoxicity. To identify potentially phototoxictetracycline derivatives, the 3T3 Neutral Red Phototoxicity assay wasemployed. The assay is described in Toxicology In Vitro 12:305-327,1998.

[0121] Briefly, 3T3 cells are seeded in to 96-well plates and incubatedover night. The growth medium is removed and replaced with phenol-redfree Hanks' Balanced Salt Solution containing serial dilutions of thetetracycline derivatives (two plates per compound). After an initial onehour incubation at 37° C., one plate is exposed to 5 Joules/cm² ofUVA/white light from a solar simulator while the other is held in thedark. The plates are then rinsed, re-fed and incubated for 24 hours.Cell visibility is measured by neutral red uptake. Phototoxicity ismeasured by the relative toxicity between the doses with and withoutlight exposure following published guidelines. (Reference compoundsinclude commercially available tetracycline, doxycycline, andminocycline.) The relative phototoxicity is called photoirritancy factor(PIF). The phototoxic response of the compounds in the present assay isconsistent with their behavior in vivo.

[0122] The class of low phototoxicity tetracyline derivatives has lessthan 75% of the phototoxicity of minocycline, preferably less than 70%,more preferably less than 60%, and most preferably 50% or less, whereinthe phototoxicity of minocycline is about 2.04. Optimally, the class oflow phototoxicity tetracycline derivatives have PIF values of 1. At aPIF value of 1, a compound is considered to have no measurablephototoxicity. Members of this class include, but are not limited to,tetracycline compounds having general forumulae:

STRUCTURE K

[0123] wherein: R7, R8, and R9 taken together in each case, have thefollowing meanings: R7 R8 R9 hydrogen hydrogen amino hydrogen hydrogenpalmitamide hydrogen hydrogen dimethylamino

STRUCTURE L STRUCTURE M

STRUCTURE N STRUCTURE O

[0124] wherein: R7, R8, and R9 taken together in each case, have thefollowing meanings: R7 R8 R9 hydrogen hydrogen acetamido hydrogenhydrogen dimethylaminoacetamido hydrogen hydrogen nitro hydrogenhydrogen amino

[0125] The class of low phototoxicity tetracycline compound derivativesincludes those derivatives having PIF values greater than 1, i.e. 1 toabout 2, preferably 1 to about 1.5. One example is a tetracyclinederivative having the general formula:

STRUCTURE K:

[0126] wherein: R7, R8, and R9 taken together are, respectively,hydrogen, hydrogen and dimethylamino.

EXAMPLES

[0127] The following examples serve to provide further appreciation ofthe invention but are not meant in any way to restrict the effectivescope of the invention.

Example 14-Dedimethylamino-7-dimethylamino-6-demethyl-6-deoxy-9-nitrotetracyclineSulfate

[0128] To a solution of one millimole of4-dedimethylamino-7-dimethylamino-6-demethyl-6-deoxytetracycline in 25ml of concentrated sulfuric acid at 0° C. was added 1.05 mmole ofpotassium nitrate. The resulting solution was stirred at ice bathtemperature for 15 minutes and poured in one liter of cold ether withstirring. The precipitated solid was allowed to settle and the majorityof solvent decanted. The remaining material was filtered through asintered glass funnel and the collected solid was washed well with coldether. The product was dried in a vacuum desiccator overnight.

Example 29-amino-4-dedimethylamino-7-dimethylamino-6-demethyl-6-deoxytetracyclineSulfate

[0129] To a solution of 300 mg of the 9-nitro compound from example 1,in 30 ml of ethanol was added 50 mg of Pt0₂. The mixture washydrogenated at atmospheric pressure until the theoretical amount ofhydrogen was absorbed. The system is flushed with nitrogen, the catalystPtO₂ is filtered and the filtrate added dropwise to 300 ml of ether. Theproduct that separates is filtered and dried in a vacuum desiccator.

Example 39-Acetamido-4-dedimethylamino-7-dimethylamino-6-demethyl-6-deoxytetracyclineSulfate

[0130] To a well stirred cold solution of 500 mg of9-amino-4-dedimethylamino-7-dimethylamino-6-demethyl-6-deoxytetracyclinesulfate from example 2, in 2.0 ml of 1.3-dimethyl-2-imidazolidinone, 500mg of sodium bicarbonate was added followed by 0.21 ml of acetylchloride. The mixture is stirred at room temperature for 30 minutes,filtered and the filtrate was added dropwise to 500 ml of ether. Theproduct that separated was filtered and dried in a vacuum desiccator.

Example 44-Dedimethylamino-7-dimethylamino-6-demethyl-6-deoxy-9-diazoniumtetracyclineSulfate

[0131] To a solution of 0.5 g of9-amino-4-dedimethylamino-7-dimethylamino-6-demethyl-6-deoxytetracyclinesulfate, from example 2, in 10 ml of 0.1N hydrochloric acid in methanolcooled in an ice bath, 0.5 ml of n-butyl nitrite was added. The solutionwas stirred at ice bath temperature for 30 minutes and then poured into250 ml of ether. The product that separated was filtered, washed withether and dried in a vacuum desiccator.

Example 59-Azido-4-dedimethylamino-7-dimethylamino-6-demethyl-6-deoxytetracyclineSulfate

[0132] To a solution of 0.3 mmole of4-dedimethylamino-7-dimethylamino-6-demethyl-6-deoxy-9-diazoniumtetracyclinesulfate, from example 4, 10 ml of 0.1 N methanolic hydrogen chloride wasadded 0.33 mmole of sodium azide. The mixture was stirred at roomtemperature for 1.5 hours. The reaction mixture was then poured into 200ml of ether. The product that separated was filtered and dried in avacuum desiccator.

Example 69-Amino-8-chloro-4-dedimethylamino-7-dimethylamino-6-demethyl-6-deoxy-tetracyclineSulfate

[0133] One gram of9-azido-4-dedimethylamino-7-dimethylamino-6-demethyl-6-deoxytetracyclinehydrochloride, from example 4, was dissolved in 10 ml of concentratedsulfuric acid saturated with HCL at 0° C. The mixture was stirred at icebath temperature for 1.5 hours and then slowly added dropwise to 500 mlof cold ether. The product that separated was filtered, washed withether and dried in a vacuum desiccator.

Example 74-Dedimethylamino-7-dimethylamino-6-demethyl-6-deoxy-9-ethoxythiocarbonylthio-tetracyclineSulfate

[0134] A solution of 1.0 mmole of4-dedimethylamino-7-dimethylamino-6-demethyl-6-deoxy-9-diazoniumtetracyclinesulfate, from example 4, in 15 ml of water was added to a solution of1.15 mmole of potassium ethyl xanthate in 15 ml of water. The mixturewas stirred at room temperature for one hour. The product separated andwas filtered and dried in a vacuum desiccator.

Example 8A General Procedure for Nitration

[0135] To 1 mmole of a 4-dedimethylamino-6-deoxytetracycline in 25 ml ofconcentrated sulfuric acid at 0° C. was added 1 mmole of potassiumnitrate with stirring. The reaction solution was stirred for 15 minutesand then poured into 100 g of chopped ice. The aqueous solution wasextracted 5 times with 20 ml of butanol each time. The butanol extractswere washed three times with 10 ml of water each time, and concentratedin vacuo to a volume of 25 ml. The light yellow crystalline solid whichprecipitated was filtered, washed with 2 ml of butanol and dried invacuo at 60° C. for 2 hours. This solid was a mixture of the twomononitro isomers.

Example 8B 4-Dedimethylamino-6-deoxy-9-nitrotetracycline

[0136] To 980 mg of the nitration product from4-dedimethylamino-6-deoxytetracycline (a mixture of the 2 isomers) in 25ml of methanol was added enough triethylamine to dissolve the solid. Thefiltered solution (pH 9.0) was adjusted to pH 5.2 with concentratedsulfuric acid. A crystalline yellow solid (236 mg.) was obtained (29%yield). The material at this point was quite pure and contained onlysmall amounts of the 7-isomer. Final purification was accomplished byliquid partition chromatography using a diatomaceous earth packed columnand the solvent system: chloroform: butanol: 0.5 M phosphate buffer (pH2) (16:1:10).

Example 9 4-Dedimethylamino-6-deoxy-7-nitrotetracycline

[0137] The methanol filtrate from example 8 was immediately adjusted topH 1.0 with concentrated sulfuric acid. The light yellow crystallinesolid, which was obtained as the sulfate salt. A purified free base wasobtained by adjusting an aqueous solution of the sulfate salt (25 mg/ml)to pH 5.2 with 2 N sodium carbonate.

Example 10 9-Amino-4-dedimethylamino-6-deoxytetracycline

[0138] To a solution of 300 mg of the 9-nitro compound, prepared inexample 8, in 30 ml of ethanol was added 50 mg of PtO₂. The mixture washydrogenated at atmospheric pressure until the theoretical amount ofhydrogen was absorbed. The system is flushed with nitrogen, the PtO₂catalyst is filtered and the filtrate added dropwise to 300 ml of ether.The solid that separates is filtered and dried in a vacuum desiccator.

Example 11 9-Acetamido-4-dedimethylamino-6-deoxytetracycline Sulfate

[0139] To well stirred cold solution of 500 mg of9-amino-4-dedimethylamino-6-deoxytetracycline sulfate, from example 10,in 2.0 ml of 1,3-dimethyl-2-imidazolidinone was added 500 mg of sodiumbicarbonate followed by 0.21 ml of acetyl chloride. The mixture wasstirred at room temperature for 30 minutes, filtered and the filtratewas added dropwise to 500 ml of ether. The solid that separated wasfiltered and dried in a vacuum desiccator.

Example 12 4-Dedimethylamino-6-deoxy-9-diazoniumtetracycline Sulfate

[0140] To a solution of 0.5 g of9-amino-4-dedimethylamino-6-deoxytetracycline sulfate, from example 10,in 10 ml of 0.1N hydrochloric acid in methanol cooled in an ice bath wasadded 0.5 ml of n-butyl nitrite. The solution was stirred at ice bathtemperature for 30 minutes and the poured into 250 ml of ether. Thesolid that separated was filtered, washed with ether and dried in avacuum desiccator.

Example 13 9-Azido-4-dedimethylamino-6-deoxytetracycline Sulfate

[0141] To a solution of 0.3 mmole of4-dedimethylamino-6-deoxy-9-diazoniumtetracycline sulfate, of example12, 10 ml of 0.1 N methanolic hydrogen chloride was added 0.33 mmole ofsodium azide. The mixture was stirred at room temperature for 1.5 hours.The reaction mixture was then poured into 200 ml of ether. The solidthat separated was filtered and dried in a vacuum desiccator.

Example 14 9-Amino-8-chloro-4-dedimethylamino-6-deoxytetracyclineSulfate

[0142] One gram of9-azido-4-dedimethylamino-7-dimethylamino-6-deoxytetracyclinehydrochloride, from example 13, was dissolved in 10 ml of concentratedsulfuric acid saturated with HCL at 0° C. The mixture was stirred at icebath temperature for 1.5 hours and then slowly added dropwise to 500 mlof cold ether. The solid that separated was filtered, washed and etherand dried in a vacuum desiccator.

Example 154-Dedimethylamino-6-deoxy-9-ethoxythiocarbonylthiotetracycline Sulfate

[0143] A solution of 1.0 mmole of4-dedimethylamino-6-deoxy-9-diazoniumtetracycline sulfate, from example12, in 15 ml of water was added to a solution of 1.15 mmole of potassiumethyl xanthate in 15 ml of water. The mixture was stirred at roomtemperature for one hour. The solid that separated was filtered anddried in a vacuum desiccator.

Example 16 9-Dimethylamino-4-dedimethylamino-6-deoxytetracycline Sulfate

[0144] To a solution of 100 mg. of the 9-amino compound from example 10,in 10 ml of ethylene glycol monomethyl ether is added 0.05 ml ofconcentrated sulfuric acid, 0.4 ml. of a 40% aqueous formaldehydesolution and 100 mg of a 10% palladium on carbon catalyst. The mixtureis hydrogenated under atmospheric pressure and room temperature for 20minutes. The catalyst was filtered and the filtrate was evaporated todryness under reduced pressure. The residue is dissolved in 5 ml ofmethanol and this solution was added to 100 ml of ether. The productthat separated was filtered and dried, yield, 98 mg.

Example 17 7-Amino-4-dedimethylamino-6-deoxytetracycline

[0145] This compound can be made using Procedure A or B. Procedure A. Toa solution of 300 mg of the 7-nitro compound, from example 1, in 30 mlof ethanol was added 50 mg of PtO₂. The mixture was hydrogenated atatmospheric pressure until the theoretical amount of hydrogen wasabsorbed. The system is flushed with nitrogen, the catalyst PtO₂ isfiltered and the filtrate added dropwise to 300 ml of ether. The solidthat separates is filtered and dried in a vacuum desiccator.

[0146] Procedure B.

[0147] 1 g of 6-deoxy-4-dedimethylamino-tetracycline was dissolved in7.6 ml THF and 10.4 ml methanesulfonic acid at −10° C. After warming themixture to 0° C. a solution of 0.86 g of dibenzyl azodicarboxylate wasadded and the mixture stirred for 2 hours at 0° C. to yield7-[1,2-bis(carbobenzyloxy)hydrazino]-4-dedimethylamino-6-deoxytetracycline.A solution of 1 millimole of this material in 70 ml 2-methoxyethanol,and 300 mg 10% Pd-C was hydrogenated at room temperature to give7-amino-6-deoxy-4-dedimethylaminotetracycline.

Example 18 7-Amino-6-deoxy-5-hydroxy-4-dedimethylaminotetracycline

[0148] 1 g of 6-deoxy-5-hydroxy-4-dedimethylaminotetracycline 3 wasdissolved in 7.6 ml THF and 10.4 ml methanesulfonic acid at −10° C.After warming the mixture to 0° C. a solution of 0.86 g dibenzylazodicarboxylate in 0.5 ml THF was added and the mixture stirred for 2hours at 0° C. to yield7-[1,2-bis(carbobenzyloxy)hydrazino]-4-dedimethylamino-6-deoxy-5-hydroxytetracycline.A solution of 1 millimole of this material in 70 ml 2-methoxyethanol,and 300 mg 10% Pd-C was hydrogenated at room temperature to give7-amino-6-deoxy-5-hydroxytetracycline.

Example 19 7-Acetamido-4-dedimethylamino-6-deoxy-5-hydroxytetracyclineSulfate

[0149] To well stirred cold solution of 500 mg of7-amino-4-dedimethylamino-6-deoxy-5-hydroxytetracycline sulfate, fromexample 18, in 2.0 ml of 1,3-dimethyl-2-imidazolidinone was added 500 mgof sodium bicarbonate followed by 0.21 ml of acetyl chloride. Themixture was stirred at room temperature for 30 minutes, filtered and thefiltrate was added dropwise to 500 ml of ether. The solid that separatedwas filtered and dried in a vacuum desiccator.

Example 20 4-Dedimethylamino-6-deoxy-5-hydroxy-7-diazoniumtetracyclineHydrochloride

[0150] To a solution of 0.5 g of7-amino-4-dedimethylamino-6-deoxy-5-hydroxytetracycline sulfate, fromexample 20, in 10 ml of 0.1N hydrochloric acid in methanol cooled in anice bath was added 0.5 ml of n-butyl nitrite. The solution was stirredat ice bath temperature for 30 minutes and then poured into 250 ml ofether. The solid that separated was filtered, washed with ether anddried in a vacuum desiccator.

Example 21 7-Azido-4-dedimethylamino-6-deoxy-5-hydroxytetracycline

[0151] To a solution of 0.3 mmole of4-dedimethylamino-6-deoxy-5-hydroxy-7-diazoniumtetracyclinehydrochloride, from example 20, 10 ml of 0.1 N methanolic hydrogenchloride was added 0.33 mmole of sodium azide. The mixture was stirredat room temperature for 1.5 hours. The reaction mixture was then pouredinto 200 ml of ether. The solid that separated was filtered and dried ina vacuum desiccator.

Example 227-Amino-8-chloro-4-dedimethylamino-6-deoxy-5-hydroxytetracycline Sulfate

[0152] One gram of7-azido-4-dedimethylamino-7-dimethylamino-6-deoxy-5-hydroxytetracyclinesulfate, from example 21, was dissolved in 10 ml of concentratedsulfuric acid (previously saturated with hydrogen chloride) at 0° C. Themixture was stirred at ice bath temperature for 1.5 hours and thenslowly added dropwise to 500 ml of cold ether. The solid that separatedwas filtered, washed with ether and dried in a vacuum desiccator.

Example 234-Dedimethylamino-6-deoxy-5-hydroxy-7-ethoxythiocarbonylthiotetracycline

[0153] A solution of 1.0 mmole of4-dedimethylamino-6-deoxy-5-hydroxy-7-diazoniumtetracyclinehydrochloride, from example 20, in 15 ml of water was added to asolution of 1.15 mmole of potassium ethyl xanthate in 15 ml of water.The mixture was stirred at room temperature for one hour. The solid thatseparated was filtered and dried in a vacuum desiccator.

Example 247-Dimethylamino-4-dedimethylamino-6-deoxy-5-hydroxytetracycline Sulfate

[0154] To a solution of 100 mg of the 7-amino compound in 10 ml ofethylene glycol monomethyl ether is added 0.05 ml of concentratedsulfuric acid, 0.4 ml of a 40% aqueous formaldehyde solution and 100 mgof a 10% palladium on carbon catalyst. The mixture is reduced withhydrogen at atmospheric pressure and room temperature for 20 minutes.The catalyst was filtered and the filtrate was evaporated to drynessunder reduced pressure. The residue is dissolved in 5 ml of methanol andthis solution was added to 100 ml of ether. The product that separatedwas filtered and dried, yield, 78 mg.

Example 25 7-Diethylamino-4-dedimethylamino-5-hydroxytetracyclineSulfate

[0155] To a solution of 100 mg of the 7-amino compound in 10 ml ofethylene glycol monomethyl ether is added 0.05 ml of concentratedsulfuric acid, 0.4 ml of acetaldehyde and 100 mg of a 10% palladium oncarbon catalyst. The mixture is reduced with hydrogen at atmosphericpressure at room temperature for 20 minutes. The catalyst was filteredand filtrate was evaporated to dryness under reduced pressure. Theresidue is dissolved in 5 ml of methanol and this solution was added to100 ml of ether. The product that separated was filtered and dried.

Example 26 4-Dedimethylamino-6-deoxy-7-diazoniumtetracyclineHydrochloride

[0156] To a solution of 0.5 g. of7-amino-4-dedimethylamino-6-deoxytetracycline sulfate, from example 17,in 10 ml of 0.1N hydrochloric acid in methanol cooled in an ice bath wasadded 0.5 ml of n-butyl nitrite. The solution was stirred at ice bathtemperature for 30 minutes and then poured into 250 ml of ether. Thesolid that separated was filtered, washed with ether and dried in avacuum desiccator.

Example 27 7-Azido-4-dedimethylamino-6-deoxytetracycline

[0157] To a solution of 0.3 mmole of4-dedimethylamino-6-deoxy-7-diazoniumtetracycline hydrochloride, fromexample 26, 10 ml of 0.1 N methanolic hydrogen chloride was added 0.33mmole of sodium azide. The mixture was stirred at room temperature for1.5 hours. The reaction mixture was then poured into 200 ml of ether.The solid that separated was filtered and dried in a vacuum desiccator.

Example 28 7-Amino-8-chloro-4-dedimethylamino-6-deoxytetracyclineSulfate

[0158] One gram of7-azido-4-dedimethylamino-7-dimethylamino-6-deoxytetracycline sulfatewas dissolved in 10 ml of concentrated sulfuric acid (previouslysaturated with hydrogen chloride) at 0° C. The mixture was stirred atice bath temperature for 1.5 hours and then slowly added dropwise to 500ml of cold ether. The solid that separated was filtered, washed withether and dried in a vacuum desiccator.

Example 294-Dedimethylamino-6-deoxy-7-ethoxythiocarbonylthiotetracycline

[0159] A solution of 1.0 mmole of4-dedimethylamino-6-deoxy-7-diazoniumtetracycline hydrochloride, fromexample 26, in 15 ml of water was added to a solution of 1.15 mmole ofpotassium ethyl xanthate in 15 ml of water. The mixture was stirred atroom temperature for one hour. The solid that separated was filtered anddried in a vacuum desiccator.

Example 30 7-Dimethylamino-4-dedimethylamino-6-deoxytetracycline Sulfate

[0160] To a solution of 100 mg of the 7-amino compound, from example 26,in 10 ml of ethylene glycol monomethyl ether is added 0.05 ml ofconcentrated sulfuric acid, 0.4 ml of a 40% aqueous formaldehydesolution and 100 mg of a 10% palladium on carbon catalyst. The mixtureis reduced with hydrogen at atmospheric pressure and room temperaturefor 20 minutes. The catalyst was filtered and the filtrate wasevaporated to dryness under reduced pressure. The residue is dissolvedin 5 ml of methanol and this solution was added to 100 ml of ether. Theproduct that separated was filtered and dried.

Example 319-Acetamido-8-chloro-4-dedimethylamino-7-dimethylamino-6-deoxy-6-demethyltetracycline

[0161] To well stirred cold solution of 500 mg of9-amino-8-chloro-4-dedimethylamino-6-deoxy-6-demethyl-7-dimethyl aminotetracycline sulfate, from example 6, in 2.0 ml of 1,3-dimethyl-2-imidazolidinone was added 500 mg of sodium bicarbonate followed by0.21 ml. of acetyl chloride. The mixture was stirred at room temperaturefor 30 minutes, filtered and the filtrate was added dropwise to 500 mlof ether. The solid that separated was filtered and dried in a vacuumdesiccator.

Example 328-Chloro-4-dedimethylamino-7-dimethylamino-6-deoxy-6-demethyl-9-ethoxythiocarbonylthiotetracycline

[0162] A solution of 1.0 mmole of-8-chloro-4-dedimethylamino-6-deoxy-6-demethyl-7-dimethylamino-9-diazoniumtetracycline hydrochloride in 15 ml of water was addedto a solution of 1.15 mmole of potassium ethyl xanthate in 15 ml ofwater. The mixture was stirred at room temperature for one hour. Thesolid that separated was filtered and dried in a vacuum desiccator.

Example 338-Chloro-9-dimethylamino-4-dedimethylamino-7-dimethylamino-6-deoxy-6-demethytetracyclineSulfate

[0163] To a solution of 100 mg. of the 9-amino compound, from example 6,in 10 ml of ethylene glycol monomethyl ether is added 0.05 ml ofconcentrated sulfuric acid, 0.4 ml of acetaldehyde and 100 mg of a 10%palladium on carbon catalyst. The mixture is reduced with hydrogen atatmospheric pressure and room temperature for 20 minutes. The catalystwas filtered and the filtrate was evaporated to dryness under reducedpressure. The residue is dissolved in 5 ml of methanol and this solutionwas added to 100 ml of ether. The product that separated was filteredand dried.

Example 34 N-(4-methylpiperazin-1-yl)methyl-4-dedimethylamino-6-demethyl-6-deoxytetracycline

[0164] An aqueous solution of 58 mg (37%) formaldehyde (0.72 mmol) wasadded to a solution of 203 mg (0.49 mmol) of4-dedimethylamino-6-demethyl-6-deoxytetracycline in 5.0 ml ethyleneglycol dimethyl ether. The mixture was stirred at room temperature for0.5 hours. 56 mg (0.56 mmol) of 1-methylpiperazine was then added andthe resulting mixture was stirred overnight and refluxed for 20 minutes.The mixture was then cooled and a solid product was collected byfiltration. The solid product was then washed with the solvent and driedby vacuum filtration.

Example 35N-(4-methylpiperazin-1-yl)methyl-4-dedimethylamino-6-demethyl-6-deoxy-9-hexanoylaminotetracycline

[0165] An aqueous solution of 49 mg 37% formaldehyde (0.60 mmol) wasadded to a solution of 146 mg (0.30 mmol) of4-dedimethylamino-6-demethyl-6-deoxy-9-hexanoylaminotetracycline in 5.0ml ethylene glycol dimethyl ether. The mixture was stirred at roomtemperature for 0.5 hours. 60 mg (0.60 mmol) of 1-methylpiperazine wasthen added and the resulting mixture was stirred overnight and refluxedfor 20 minutes. The mixture was then cooled and a solid product wascollected by filtration. The solid product was then washed with thesolvent and dried by vacuum filtration.

Example 364-Dedimethylamino-6-demethyl-6-deoxy-9-hexanoylaminotetracycline

[0166] 1.54 g (7.2 mmol) of hexanoic anhydride and 150 mg of 10% Pd/Ccatalyst were added to 300 mg (0.72 mmol) of4-dedimethylamino-6-demethyl-6-deoxytetracycline in 6.0 ml of1,4-dioxane and 6.0 ml of methanol. The mixture was hydrogenatedovernight at room temperature. The catalyst was removed by filtrationand the filtrate was concentrated under reduced pressure. The residuewas dissolved in 7 ml of ethyl acetate and trituated with 50 ml ofhexane to produce a solid product. The solid product was filtered anddried by vacuum filtration.

[0167] Thus, while there have been described what are presently believedto be the preferred embodiments of the present invention, those skilledin the art will realize that other and further embodiments can be madewithout departing from the spirit of the invention, and it is intendedto include all such further modifications and changes as come within thetrue scope of the claims set forth herein.

Example 37 Phototoxicity Determination

[0168] BALB/c 3T3 (CCL-163) cells were obtained from ATCC and culturedin antibiotic-free Dulbecco's Minimum Essential Medium (4.5 g/lglucose)(DMEM) supplemented with L-glutamine (4 mM) and 10% newborn calfserum. The working cell bank was prepared and found to be free ofmycoplasma. Streptomycin sulfate (100 μg/ml) and penicillin (100 IU/ml)were added to the medium after the cells were treated with test articlein 96-well plates.

[0169] Serial dilutions of the tetracycline derivatives were prepared inDMSO at concentrations 100× to final testing concentration. The CMTdilutions in DMSO were then diluted in Hanks' Balanced Salt Solution(HBSS) for application to the cells. The final DMSO concentration was 1%in treated and control cultures. For the dose range finding assay, 8serial dilutions covered a range of 100 to 0.03 mg/ml in half log stepswhile the definitive assays used 6 to 8 doses prepared in quarter logsteps, centered on the expected 50% toxicity point. In many cases, thedose range for treatment without UV light was different from the doserange selected with UV light. One hundred μg/ml is the highest doserecommended to prevent false negative results from UV absorption by thedosing solutions.

[0170] Controls:

[0171] Each assay included both negative (solvent) and positivecontrols. Twelve wells of negative control cultures were used on each96-well plate. Chlorpromazine (Sigma) was used as the positive controland was prepared and dosed like the test tetracycline derivatives.

[0172] Solar Simulator:

[0173] A Dermalight SOL 3 solar simulator, equipped with a UVA H1 filter(320-400 nm), was adjusted to the appropriate height. Measurement ofenergy through the lid of a 96-well microtiter plate was carried outusing a calibrated UV radiometer UVA sensor. Simulator height wasadjusted to deliver 1.7±0.1 m/Wcm² of UVA energy (resulting dose was1J/cm² per 10 min.)

[0174] Phototoxicity Assay:

[0175] Duplicate plates were prepared for each test material by seeding10⁴ 3T3 cells per well in μl of complete medium 24 hours beforetreatment. Prior to treatment, the medium was removed, and the cellswashed once with 125 μl prewarmed HBSS. Fifty μl of prewarmed HBSS wereadded to each well. Fifty μl of test article dilutions were added to theappropriate wells and the plates returned to the incubator forapproximately one hour. Following the 1 hr incubation, the platesdesignated for the photoirritation assay were exposed (with the lid on)to 1.7±0.1 mW/cm² UVA light for 50±2 minutes at room temperatureresulting in an irradiation dose of 5J/cm2. Duplicate plates designatedfor the cytotoxicity assay were kept in the dark room temperature for50±2 minutes. After the 50 minute exposure period the test articledilutions were decanted from the plates and the cells washed once with125 μl HBSS. One hundred μl of medium were added to all wells and thecells incubated as above for 24±1 hours.

[0176] After 24 hours of incubation, the medium was decanted and 100 μlof the Neutral Red containing media added to each well. The plates werereturned to the incubator and incubated for approximately 3 hours. After3 hours, the medium was decanted and each well rinsed once with 250 μlof HBSS. The plates were blotted to remove the HBSS and 100 μl ofNeutral Red Solvent were added to each well. After a minimum of 20minutes of incubation at room temperature (with shaking), the absorbanceat 550 nm was measured with a plate reader, using the mean of the blankouter wells as the reference. Relative survival was obtained bycomparing the amount of neutral red taken by test article and positivecontrol treated groups to the neutral red taken up by the negative groupon the same plate. IC₅₀ values for both the UVA exposed and non-exposedgroups were determined whenever possible. One dose range finding and atleast two definitive trails were performed on each tetracyclinederivative and control compound.

[0177] Determination of Phototoxicity:

[0178] Phototoxicity of the tetracycline derivatives can be measured byits photoirritancy factor (PIF). The PIF was determined by comparing theIC₅₀ without UVA [IC₅₀(−UVA)] with the IC₅₀ with UVA [IC o(+UVA)]:${PIF} = \frac{{IC}_{50}\left( {- {UVA}} \right)}{{IC}_{50}\left( {+ {UVA}} \right)}$

[0179] If both IC₅₀ values can be determined, the cut off value of thefactor to discriminate between phototoxicants and non-phototoxicants isa factor of 5. A factor greater than 5 is indicative of phototoxicpotential of the test material.

[0180] If IC₅₀ (+UVA) can be determined but IC₅₀(−UVA) cannot, the PIFcannot be calculated, although the compound tested may have some levelof phototoxic potential. In this case, a “>PIF” can be calculated andthe highest testable dose (−UVA) will be used for calculation of the“>PIF.”${> {PIF}} = \frac{{maximum}\quad {{dose}\left( {- {UVA}} \right)}}{{IC}_{50}\left( {+ {UVA}} \right)}$

[0181] If both, IC₅₀(+UVA) and IC₅₀(+UVA) cannot be calculated becausethe chemical does not show cytotoxicty (50% reduction in viability) upto the highest dose tested, this would indicate a lack of phototoxicpotential.

What is claimed is:
 1. A tetracycline compound of the formulae:

wherein: R7 is selected from the group consisting of hydrogen, amino,nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of hydrogen, amino,azido, nitro, acylamino, hydroxy, ethoxythiocarbonylthio, mono(loweralkyl) amino, halogen, diazonium, di(lower alkyl)amino and RCH(NH₂)CO; Ris hydrogen or lower alkyl; and pharmaceutically acceptable andunacceptable salts thereof; with the following provisos: when either R7and R9 are hydrogen then R8 must be halogen; and when R6-a, R6, R5 andR9 are all hydrogen and R7 is hydrogen, amino, nitro, halogen,dimethylamino or diethylamino, then R8 must be halogen; and when R6-a ismethyl, R6 and R9 are both hydrogen, R5 is hydroxyl, and R7 is hydrogen,amino, nitro, halogen or diethylamino, then R8 is halogen; and when R6-ais methyl, R6 is hydroxyl, R5, R7 and R9 are all hydrogen, then R8 mustbe halogen; and when R6-a, R6 and R5 are all hydrogen, R9 is methylaminoand R7 is dimethylamino, then R8 must be halogen; and when R6-a ismethyl, R6 is hydrogen, R5 is hydroxyl, R9 is methylamino and R7 isdimethylamino, then R8 must be halogen; and when R6-a is methyl, R6, R5and R9 are all hydrogen and R7 is cyano, then R8 must be halogen.
 2. Atetracycline compound of the formulae:

wherein: R7 is selected from the group consisting of hydrogen, amino,nitro, mono(lower alkyl) amino, halogen, and di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R4 is selected from the group consisting of NOH, N-NH-A, andNH-A, where A is a lower alkyl group; R8 is selected from the groupconsisting of hydrogen and halogen; R9 is selected from the groupconsisting of hydrogen, amino, azido, nitro, acylamino, hydroxy,ethoxythiocarbonylthio, mono(lower alkyl) amino, halogen, di(loweralkyl)amino and RCH(NH₂)CO; R is hydrogen or lower alkyl; andpharmaceutically acceptable and unacceptable salts thereof; with thefollowing provisos: when R4 is NOH, N-NH-alkyl or NH-alkyl and R7, R6-a,R6, R5, and R9 are all hydrogen, then R8 must be halogen; and when R4 isNOH, R6-a is methyl, R6 is hydrogen or hydroxyl, R7 is halogen, R5 andR9 are both hydrogen, then R8 must be halogen; and when R4 isN-NH-alkyl, R6-a is methyl, R6 is hydroxyl and R7, R5, R9 are allhydrogen, then R8 must be halogen; and when R4 is NH-alkyl, R6-a, R6, R5and R9 are all hydrogen, R7 is hydrogen, amino, mono(lower alkyl)amino,halogen, di(lower alkyl)amino or hydroxyl, then R8 must be halogen; andwhen R4 is NH-alkyl, R6-a is methyl, R6 and R9 are both hydrogen, R5 ishydroxyl, and R7 is mono(lower alkyl)amino or di(lower alkyl)amino, thenR8 must be halogen; and when R4 is NH-alkyl, R6-a is methyl, R6 ishydroxy or hydrogen and R7, R5, and R9 are all be hydrogen, then R8 mustbe halogen.
 3. A 4-dedimethylamino tetracycline compound selected from:

wherein: R7, R8, and R9 taken together in each case, have the followingmeanings: R7 R8 R9 azido hydrogen hydrogen dimethylamino hydrogen azidohydrogen hydrogen amino hydrogen hydrogen azido hydrogen hydrogen nitrodimethylamino hydrogen amino acylamino hydrogen hydrogen hydrogenhydrogen acylamino amino hydrogen nitro hydrogen hydrogen(N,N-dimethyl)glycylamino amino hydrogen amino hydrogen hydrogenethoxythiocarbonylthio dimethylamino hydrogen acylamino dimethylaminohydrogen diazonium dimethylamino chloro amino hydrogen chloro aminoamino chloro amino acylamino chloro acylamino amino chloro hydrogenacylamino chloro hydrogen monoalkylamino chloro amino nitro chloro aminodimethylamino chloro acylamino dimethylamino chloro dimethylaminohydrogen hydrogen dimethylamino dimethylamino hydrogen hydrogen and

wherein: R7, R8, and R9 taken together in each case, have the followingmeanings: R7 R8 R9 azido hydrogen hydrogen dimethylamino hydrogen azidohydrogen hydrogen amino hydrogen hydrogen azido hydrogen hydrogen nitrodimethylamino hydrogen amino acylamino hydrogen hydrogen hydrogenhydrogen acylamino amino hydrogen nitro hydrogen hydrogen(N,N-dimethyl)glycylamino amino hydrogen amino hydrogen hydrogenethoxythiocarbonylthio dimethylamino hydrogen acylamino hydrogenhydrogen diazonium hydrogen hydrogen dimethylamino diazonium hydrogenhydrogen ethoxythiocarbonylthio hydrogen hydrogen dimethylamino chloroamino amino chloro amino acylamino chloro acylamino hydrogen chloroamino amino chloro hydrogen acylamino chloro hydrogen monoalkylaminochloro amino nitro chloro amino and

wherein: R8 is hydrogen or halogen and R9 is selected from the groupconsisting of nitro, (N,N-dimethyl)glycylamino, andethoxythiocarbonylthio; and

wherein: R7, R8, and R9 taken together in each case, have the followingmeanings: R7 R8 R9 amino hydrogen hydrogen nitro hydrogen hydrogen azidohydrogen hydrogen dimethylamino hydrogen azido hydrogen hydrogen aminohydrogen hydrogen azido hydrogen hydrogen nitro bromo hydrogen hydrogendimethylamino hydrogen amino acylamino hydrogen hydrogen hydrogenhydrogen acylamino amino hydrogen nitro hydrogen hydrogen(N,N-dimethyl)glycylamino amino hydrogen amino diethylamino hydrogenhydrogen hydrogen hydrogen ethoxythiocarbonylthio dimethylamino hydrogenmethylamino dimethylamino hydrogen acylamino dimethylamino chloro aminoamino chloro amino acylamino chloro acylamino hydrogen chloro aminoamino chloro hydrogen acylamino chloro hydrogen monoalkylamino chloroamino nitro chloro amino and pharmaceutically acceptable andunacceptable salts thereof.


4. A tetracycline compound of the formulae:

wherein: R7 is selected from the group consisting of hydrogen, amino,nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of hydrogen, amino,azido, nitro, acylamino, hydroxy, ethoxythiocarbonylthio, mono(loweralkyl) amino, halogen, diazonium, di(lower alkyl)amino and RCH(NH₂)CO; Ris hydrogen or lower alkyl; R^(a) and R^(b) are selected from the groupconsisting of hydrogen, methyl, ethyl, n-propyl and 1-methylethyl withthe proviso that R^(a) and R^(b) cannot both be hydrogen; R^(c) andR^(d) are, independently, (CH₂)_(n)CHR^(e) wherein n is 0 or 1 and R^(e)is selected from the group consisting of hydrogen, alkyl, hydroxy,lower(C₁-C₃) alkoxy, amino, or nitro; and, W is selected from the groupconsisting of (CHR^(e))_(m) wherein m is 0-3 and said R^(e) is as above,NH, N(C₁-C₃) straight chained or branched alkyl, O, S and N(C₁-C₄)straight chain or branched alkoxy; and, pharmaceutically acceptable andunacceptable salts thereof.
 5. A tetracycline compound according toclaim 4 with the following provisos: when either R7 and R9 are hydrogenthen R8 must be halogen; and when R6-a, R6, R5 and R9 are all hydrogenand R7 is hydrogen, amino, nitro, halogen, dimethylamino ordiethylamino, then R8 must be halogen; and when R6-a is methyl, R6 andR9 are both hydrogen, R5 is hydroxyl, and R7 is hydrogen, amino, nitro,halogen or diethylamino, then R8 is halogen; and when R6-a is methyl, R6is hydroxyl, R5, R7 and R9 are all hydrogen, then R8 must be halogen;and when R6-a, R6 and R5 are all hydrogen, R9 is methylamino and R7 isdimethylamino, then R8 must be halogen; and when R6-a is methyl, R6 ishydrogen, R5 is hydroxyl, R9 is methylamino and R7 is dimethylamino,then R8 must be halogen; and when R6-a is methyl, R6, R5 and R9 are allhydrogen and R7 is cyano, then R8 must be halogen.
 6. A tetracyclinecompound according to claim 1 wherein the PIF is about
 1. 7. Atetracycline compound according to claim 6 selected from the groupconsisting of: STRUCTURE K wherein: R7, R8, and R9 taken together ineach case, have the following meanings: R7 R8 R9 hydrogen hydrogen aminohydrogen hydrogen palmitamide

and STRUCTURE L STRUCTURE M STRUCTURE N STRUCTURE O wherein: R7, R8, andR9 taken together in each case, have the following meanings: R7 R8 R9hydrogen hydrogen acetamido hydrogen hydrogen dimethylaminoacetamidohydrogen hydrogen nitro hydrogen hydrogen amino

and STRUCTURE P wherein: R7, R8, and R9 taken together are,respectively, hydrogen, hydrogen and nitro.
 8. A tetracycline compoundaccording to claim I wherein the PIF has a value between 1 and
 2. 9. Atetracycline compound according to claim 8 having general forumula:STRUCTURE K wherein: R7, R8, and R9 taken together are, respectively,hydrogen, hydrogen and dimethylamino.
 10. A tetracycline compound of theformulae: STRUCTURE C STRUCTURE D STRUCTURE E STRUCTURE F wherein: R7 isselected from the group consisting of an aryl, alkenyl and alkynyl; R6-ais selected from the group consisting of hydrogen and methyl; R6 and R5are selected from the group consisting of hydrogen and hydroxyl; R8 isselected from the group consisting of hydrogen and halogen; R9 isselected from the group consisting of hydrogen, amino, azido, nitro,acylamino, hydroxy, ethoxythiocarbonylthio, mono(lower alkyl) amino,halogen, diazonium, di(lower alkyl)amino and RCH(NH₂)CO; andpharmaceutically acceptable and unacceptable salts thereof; or STRUCTUREC STRUCTURE D STRUCTURE E STRUCTURE F wherein: R7 is selected from thegroup consisting of hydrogen, amino, nitro, mono(lower alkyl) amino,halogen, di(lower alkyl)amino, ethoxythiocarbonylthio, azido, acylamino,diazonium, cyano, and hydroxyl; R6-a is selected from the groupconsisting of hydrogen and methyl; R6 and R5 are selected from the groupconsisting of hydrogen and hydroxyl; R8 is selected from the groupconsisting of hydrogen and halogen; R9 is selected from the groupconsisting of an aryl, alkenyl and alkynyl; and pharmaceuticallyacceptable and unacceptable salts thereof; or STRUCTURE C STRUCTURE DSTRUCTURE E STRUCTURE F wherein: R7 and R9 are selected from the groupconsisting of an aryl, alkene alkyne, or mixures thereof; R6-a isselected from the group consisting of hydrogen and methyl; R6 and R5 areselected from the group consisting of hydrogen and hydroxyl; R8 isselected from the group consisting of hydrogen and halogen; andpharmaceutically acceptable and unacceptable salts thereof.
 11. Atetracycline compound of the formulae: STRUCTURE G STRUCTURE H STRUCTUREI STRUCTURE J wherein: R7 is selected from the group consisting of anaryl, alkenyl and alkynyl; R6-a is selected from the group consisting ofhydrogen and methyl; R6 and R5 are selected from the group consisting ofhydrogen and hydroxyl; R4 is selected from the group consisting of NOH,N-NH-A, and NH-A, where A is a lower alkyl group; R8 is selected fromthe group consisting of hydrogen and halogen; R9 is selected from thegroup consisting of hydrogen, amino, azido, nitro, acylamino, hydroxy,ethoxythiocarbonylthio, mono(lower alkyl) amino, halogen, di(loweralkyl)amino and RCH(NH₂)CO; and pharmaceutically acceptable andunacceptable salts thereof; or STRUCTURE G STRUCTURE H STRUCTURE ISTRUCTURE J wherein: R7 is selected from the group consisting ofhydrogen, amino, nitro, mono(lower alkyl) amino, halogen, di(loweralkyl)amino, ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano,and hydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R4 is selected from the group consisting of NOH, N-NH-A, andNH-A, where A is a lower alkyl group; R8 is selected from the groupconsisting of hydrogen and halogen; R9 is selected from the groupconsisting of an aryl, alkenyl and alkynyl; and pharmaceuticallyacceptable and unacceptable salts thereof; or STRUCTURE G STRUCTURE HSTRUCTURE I STRUCTURE J wherein: R7 and R9 are selected from the groupconsisting of an aryl, alkenyl, alkynyl; or mixtures thereof; R6-a isselected from the group consisting of hydrogen and methyl; R6 and R5 areselected from the group consisting of hydrogen and hydroxyl; R4 isselected from the group consisting of NOH, N-NH-A, and NH-A, where A isa lower alkyl group; and R8 is selected from the group consisting ofhydrogen and halogen; and pharmaceutically acceptable and unacceptablesalts thereof;
 12. A tetracycline compound of the formulae: STRUCTURE Kwherein: R7 is selected from the group consisting of an aryl, alkenyland alkynyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of hydrogen, amino,azido, nitro, acylamino, hydroxy, ethoxythiocarbonylthio, mono(loweralkyl) amino, halogen, di(lower alkyl)amino and RCH(NH₂)CO; andpharmaceutically acceptable and unacceptable salts thereof, or STRUCTUREK wherein: R7 is selected from the group consisting of hydrogen, amino,nitro, mono(lower alkyl) amino, halogen, di(lower alkyl)amino,ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano, andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of an aryl, alkenyland alkynyl; and pharmaceutically acceptable and unacceptable saltsthereof, or STRUCTURE K wherein: R7 and R9 are selected from the groupconsisting of an aryl, alkenyl, alkynyl and mixtures thereof; and R8 isselected from the group consisting of hydrogen and halogen; andpharmaceutically acceptable and unacceptable salts thereof; andSTRUCTURE L STRUCTURE M STRUCTURE N STRUCTURE O wherein: R7 is selectedfrom the group consisting of an aryl, alkenyl and alkynyl; R8 isselected from the group consisting of hydrogen and halogen; andpharmaceutically acceptable and unacceptable salts thereof; or STRUCTUREL STRUCTURE M STRUCTURE N STRUCTURE O wherein: R7 is selected from thegroup consisting of hydrogen, amino, nitro, mono(lower alkyl) amino,halogen, di(lower alkyl)amino, ethoxythiocarbonylthio, azido, acylamino,diazonium, cyano, and hydroxyl; R8 is selected from the group consistingof hydrogen and halogen; R9 is selected from the group consisting of anaryl, alkenyl and alkynyl; and pharmaceutically acceptable andunacceptable salts thereof; or STRUCTURE L STRUCTURE M STRUCTURE NSTRUCTURE O wherein: R7 is and R9 are selected from the group consistingof an aryl, alkenyl, alkynyl and mixtures thereof; R8 is selected fromthe group consisting of hydrogen and halogen; R9 is selected from thegroup consisting of hydrogen, amino, azido, nitro, acylamino, hydroxy,ethoxythiocarbonylthio, mono(lower alkyl) amino, halogen, di(loweralkyl)amino and RCH(NH₂)CO; and pharmaceutically acceptable andunacceptable salts thereof; and STRUCTURE P wherein: R9 is selected fromthe group consisting of an aryl, alkenyl and alkynyl; and R8 is selectedfrom the group consisting of hydrogen and halogen; and pharmaceuticallyacceptable and unacceptable salts thereof; and STRUCTURE Q STRUCTURE Rwherein: R7 is selected from the group consisting of an aryl, alkenyland alkynyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of hydrogen, amino,azido, nitro, acylamino, hydroxy, ethoxythiocarbonylthio, mono(loweralkyl) amino, halogen, di(lower alkyl)amino and RCH(NH₂)CO; andpharmaceutically acceptable and unacceptable salts thereof; or STRUCTUREQ STRUCTURE R wherein: R7 is selected from the group consisting ofhydrogen, amino, nitro, mono(lower alkyl) amino, halogen, di(loweralkyl)amino, ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano,and hydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of an aryl, alkenyland alkynyl; and pharmaceutically acceptable and unacceptable saltsthereof; or STRUCTURE Q STRUCTURE R wherein: R7 and R9 are selected fromthe group consisting of an aryl, alkenyl, alkynyl; and mixtures thereof;R8 is selected from the group consisting of hydrogen and halogen; andpharmaceutically acceptable and unacceptable salts thereof.
 13. Atetracycline compound of the formulae: STUCTURES S-Z wherein: R7 isselected from the group consisting of an aryl, alkenyl and alkynyl; R6-ais selected from the group consisting of hydrogen and methyl; R6 and R5are selected from the group consisting of hydrogen and hydroxyl; R8 isselected from the group consisting of hydrogen and halogen; R9 isselected from the group consisting of hydrogen, amino, azido, nitro,acylamino, hydroxy, ethoxythiocarbonylthio, mono(lower alkyl) amino,halogen, diazonium, di(lower alkyl)amino and RCH(NH₂)CO; R^(a) and R^(b)are selected from the group consisting of hydrogen, methyl, ethyl,n-propyl and 1-methylethyl with the proviso that R^(a) and R^(b) cannotboth be hydrogen; R^(c) and R^(d) are, independently, (CH₂)_(n)CHR^(e)wherein n is 0 or 1 and R^(e) is selected from the group consisting ofhydrogen, alkyl, hydroxy, lower(C₁-C₃) alkoxy, amino, or nitro; and, Wis selected from the group consisting of (CHR^(e))_(m) wherein m is 0-3and said R^(e) is as above, NH, N(C₁-C₃) straight chained or branchedalkyl, O, S and N(C₁-C₄) straight chain or branched alkoxy; and,pharmaceutically acceptable and unacceptable salts thereof; orSTRUCTURES S-Z wherein: R7 is selected from the group consisting ofhydrogen, amino, nitro, mono(lower alkyl) amino, halogen, di(loweralkyl)amino, ethoxythiocarbonylthio, azido, acylamino, diazonium, cyano,and hydroxyl; R6-a is selected from the group consisting of hydrogen andmethyl; R6 and R5 are selected from the group consisting of hydrogen andhydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R9 is selected from the group consisting of an aryl, alkenyland alkynyl; R^(a) and R^(b) are selected from the group consisting ofhydrogen, methyl, ethyl, n-propyl and 1-methylethyl with the provisothat R^(a) and R^(b) cannot both be hydrogen; R^(c) and R^(d) are,independently, (CH₂)_(n)CHR^(e) wherein n is 0 or 1 and R^(e) isselected from the group consisting of hydrogen, alkyl, hydroxy,lower(C₁-C₃) alkoxy, amino, or nitro; and, W is selected from the groupconsisting of (CHR^(e))_(m) wherein m is 0-3 and said R^(e) is as above,NH, N(C₁-C₃) straight chained or branched alkyl, O, S and N(C₁-C₄)straight chain or branched alkoxy; and pharmaceutically acceptable andunacceptable salts thereof; or STRUCTURES S-Z wherein: R7 and R9 areselected from the group consisting of an aryl, alkenyl, alkynyl andmixtures thereof; R6-a is selected from the group consisting of hydrogenand methyl; R6 and R5 are selected from the group consisting of hydrogenand hydroxyl; R8 is selected from the group consisting of hydrogen andhalogen; R^(a) and R^(b) are selected from the group consisting ofhydrogen, methyl, ethyl, n-propyl and 1-methylethyl with the provisothat R^(a) and R^(b) cannot both be hydrogen; R^(c) and R^(d) are,independently, (CH₂)_(n)CHR^(e) wherein n is 0 or 1 and R^(e) isselected from the group consisting of hydrogen, alkyl, hydroxy,lower(C₁-C₃) alkoxy, amino, or nitro; and, W is selected from the groupconsisting of (CHR^(e))_(m) wherein m is 0-3 and said R^(e) is as above,NH, N(C₁-C₃) straight chained or branched alkyl, O, S and N(C₁-C₄)straight chain or branched alkoxy; and, pharmaceutically acceptable andunacceptable salts thereof
 14. A method for treating a mammal sufferingfrom a condition that benefits from a non-antimicrobial dose of atetracycline compound, said condition being characterized by excessivecollagen destruction, excessive MMP enzyme activity, excessive TNFactivity, excessive nitric oxide activity, excessive IL-1 activity,excessive elastase activity, excessive loss of bone density, excessiveprotein degradation, excessive muscle wasting, excessive glycosylationof collagen, excessive COX-2 activity, insufficient bone proteinsynthesis, insufficient interleukin-10 production or excessivephospholipase A₂ activity the method comprising administering to themammal an effective amount of a tetracycline compound according toclaim
 1. 15. A method according to claim 10, wherein said condition isabdominal aortic aneurysm, ulceration of the cornea, periodontaldisease, diabetes, diabetes mellitus, scleroderma, progeria, lungdisease, cancer, graft versus host disease, disease of depressed bonemarrow function, thrombocytopenia, prosthetic joint loosening,spondyloarthropathies, osteoporosis, Paget's disease, autoimmunedisease, systemic lupus erythematosus, acute or chronic inflammatorycondition, renal disease or connective tissue disease.
 16. A methodaccording to claim 11, wherein said acute or chronic inflammatorycondition is inflammatory bowel disease, arthritis, osteoarthritis,rheumatoid arthritis, pancreatitis, nephritis, glomerulonephritis,sepsis, septic shock, lipopolysaccharide endotoxin shock, multisystemorgan failure or psoriasis.
 17. A method according to claim 11, whereinsaid lung disease is ARDS, cystic fibrosis, emphysema or acute lunginjury resulting from inhalation of toxicants.
 18. A method according toclaim 11, wherein said renal disease is chronic renal failure, acuterenal failure, nephritis or glomerulonephritis.
 19. A method fortreating a mammal suffering from a neurological or neurodegenerativecondition that benefits from a non-antimicrobial dose of a tetracyclinecompound, the method comprising administering to the mammal an effectiveamount of a tetracycline compound according to any of claims 1
 20. Amethod according to claim 19, wherein said condition is Alzheimer'sdisease, Guillain-Barré Syndrome, Krabbe's disease,adrenoleukodystrophy, Parkinson's disease, Huntington's disease,multiple sclerosis, amyotrophic lateral sclerosis or an encephalopathy.21. A method according to claim 16, wherein said encephalopathy is aspongiform encephalopathy.